U.S. patent application number 11/993335 was filed with the patent office on 2011-01-27 for gene expression technique.
This patent application is currently assigned to Novozymes Delta Limited. Invention is credited to Leslie Robert Evans, Christopher John Arthur Finnis, Thomas Payne, Darrell Sleep.
Application Number | 20110020865 11/993335 |
Document ID | / |
Family ID | 34855958 |
Filed Date | 2011-01-27 |
United States Patent
Application |
20110020865 |
Kind Code |
A1 |
Payne; Thomas ; et
al. |
January 27, 2011 |
Gene Expression Technique
Abstract
The present invention provides a host cell suitable for enhanced
production of a protein product of choice characterised in that the
host cell is genetically modified to cause over-expression of two
or more helper proteins selected from a DnaJ-like protein (such as
JEM1), an Hsp70 family protein (such as LHS1) and SIL1 wherein at
least one of the over-expressed two or more helper proteins is
selected from JEM1, LHS1 and SIL1, and wherein the DnaJ-like
protein is not SCJ1.
Inventors: |
Payne; Thomas; (Nottingham,
GB) ; Sleep; Darrell; (Notitngham, GB) ;
Finnis; Christopher John Arthur; (Nottingham, GB) ;
Evans; Leslie Robert; (Nottingham, GB) |
Correspondence
Address: |
NOVOZYMES NORTH AMERICA, INC.
500 FIFTH AVENUE, SUITE 1600
NEW YORK
NY
10110
US
|
Assignee: |
Novozymes Delta Limited
Nottingham
GB
University of Nottigham
Nottingham
GB
|
Family ID: |
34855958 |
Appl. No.: |
11/993335 |
Filed: |
June 22, 2006 |
PCT Filed: |
June 22, 2006 |
PCT NO: |
PCT/GB06/02289 |
371 Date: |
July 7, 2010 |
Current U.S.
Class: |
435/69.1 ;
435/254.21; 435/471 |
Current CPC
Class: |
C12N 15/81 20130101;
C12P 21/00 20130101; C07K 14/395 20130101 |
Class at
Publication: |
435/69.1 ;
435/254.21; 435/471 |
International
Class: |
C12P 21/06 20060101
C12P021/06; C12N 1/19 20060101 C12N001/19; C12N 15/74 20060101
C12N015/74 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 22, 2005 |
GB |
0512707.1 |
Claims
1-31. (canceled)
32. A host cell suitable for enhanced production of a protein
product of choice characterised in that the host cell is
genetically modified to cause over-expression of two or more helper
proteins selected from a DnaJ-like protein (such as JEM1), an Hsp70
family protein (such as LHS1) and SIL1, wherein at least one of the
over-expressed two or more helper proteins is selected from JEM1,
LHS1 and SIL1, and wherein the DnaJ-like protein is not SCJ1.
33. The host cell of claim 32 wherein the host cell is genetically
modified to cause over-expression of (a) a DnaJ-like protein and an
Hsp70 family protein; or (b) a DnaJ-like protein and SIL1; or (c)
an Hsp70 family protein and SIL1.
34. A host cell suitable for enhanced production of a protein
product of choice characterised in that the host cell is
genetically modified to cause over-expression of three or more
helper proteins, wherein the three or more helper proteins comprise
a DnaJ-like protein, an Hsp70 family protein and SIL1, and wherein
the DnaJ-like protein is not SCJ1.
35. The host cell of claim 32 wherein the Hsp70 family protein is a
protein that localises to the lumen of the ER.
36. The host cell of claim 2 wherein the Hsp70 family protein is
not a prokaryotic Hsp70 family protein.
37. The host cell of claim 32 according to any one of the preceding
claims wherein the Hsp70 family protein is LHS1, KAR2, SSA1, SSA2,
SSA3, SSA4, SSE1, SSE2, SSB1, SSB2 or ECM10.
38. The host cell of claim 32 wherein the DnaJ-like protein is a
protein that localises to the ER membrane.
39. The host cell of claim 32 wherein the DnaJ-like protein is
selected from JEM1, MDJ2, SEC63, YDJ1, XDJ1, APJ1, SIS1, DJP1,
ZUO1, SWA2, JJJ1, JJJ2, JJJ3, CAJ1, CWC23, PAM18, JAC1, JID1, SCJ1,
HLJ1 and ERJ5.
40. The host cell of claim 32 wherein the host cell is further
genetically modified to cause over-expression of at least one, two,
three, four, five, six or seven proteins involved in the formation
of disulphide bonds in other proteins selected from the group
consisting of ERO1, ERV2, EUG1, MPD1, MPD2, EPS1 and PDI1.
41. The host cell that is suitable for enhanced production of a
protein product of choice characterised in that the host cell
comprises a first gene encoding a first helper protein selected
from JEM1, LHS1 or SIL1 or a variant thereof, and a second gene
encoding a desired protein product of choice, wherein the host cell
is genetically modified to cause over-expression of the first
helper protein, and (a) wherein the first and second genes are not
both present within the host cell on the same 2 .mu.m-family
plasmid; and (b) wherein the host cell is not genetically modified
to cause over-expression of a further helper protein that is
different from the first helper protein and is selected from the
group consisting of AHA1, CCT2, CCT3, CCT4, CCT5, CCT6, CCT7, CCT8,
CNS1, CPR3, CPR6, ERO1, EUG1, FMO1, HCH1, HSP10, HSP12, HSP104,
HSP26, HSP30, HSP42, HSP60, HSP78, HSP82, JEM1, MDJ1, MDJ2, MPD1,
MPD2, PDI1, PFD1, ABC1, APJ1, ATP11, ATP12, BTT1, CDC37, CPR7,
HSC82, KAR2, LHS1, MGE1, MRS11, NOB1, ECM10, SSA1, SSA2, SSA3,
SSA4, SSC1, SSE2, SIL1, SLS1, ORM1, ORM2, PER1, PTC2, PSE1, UBI4
and HAC1 or a truncated intronless HAC1.
42. The host cell of claim 41, wherein the first helper protein is
JEM1, LHS1 or SIL1.
43. The host cell of claim 42 wherein the first helper protein is
the only helper protein that is over-expressed by the host
cell.
44. The host cell of claim 32 wherein the protein product of choice
is a heterologous protein and/or comprises a leader sequence
effective to cause secretion.
45. The host cell of claim 32 wherein the protein product of choice
is a eukaryotic protein, or a fragment or variant thereof.
46. The host cell of claim 32 wherein the protein product of choice
comprises albumin, a monoclonal antibody, an etoposide, a serum
protein, antistasin, a tick anticoagulant peptide, transferrin,
lactoferrin, endostatin, angiostatin, collagens, immunoglobulins,
or immunoglobulin-based molecules or fragment of either, a Kunitz
domain protein, interferons, interleukins, leptin, CNTF and
fragments thereof, IL1-receptor antagonist, erythropoietin (EPO)
and EPO mimics, thrombopoietin (TPO) and TPO mimics, prosaptide,
cyanovirin-N, 5-helix, T20 peptide, T1249 peptide, HIV gp41, HIV
gp120, urokinase, prourokinase, tPA, hirudin, platelet derived
growth factor, parathyroid hormone, proinsulin, insulin, glucagon,
glucagon-like peptides, insulin-like growth factor, calcitonin,
growth hormone, transforming growth factor beta, tumour necrosis
factor, G-CSF, GM-CSF, M-CSF, FGF, coagulation factors in both pre
and active forms, including but not limited to plasminogen,
fibrinogen, thrombin, pre-thrombin, pro-thrombin, von Willebrand's
factor, alpha,-antitrypsin, plasminogen activators, Factor VII,
Factor VIII, Factor IX, Factor X and Factor XIII, nerve growth
factor, LACI, platelet-derived endothelial cell growth factor
(PD-ECGF), glucose oxidase, serum cholinesterase, inter-alpha
trypsin inhibitor, antithrombin III, apo-lipoprotein species,
Protein C, Protein S, or a variant or fragment of any of the above,
or a fusion of albumin and any of the above.
47. The host cell of claim 32 wherein the protein product of choice
comprises the sequence of albumin or a variant or fragment
thereof.
48. The host cell of claim 32 wherein the protein product of choice
comprises the sequence of transferrin family member, or a variant
or fragment thereof.
49. The host cell of claim 32 wherein the protein product of choice
comprises a fusion protein.
50. The host cell of claim 32 comprising an exogenous
polynucleotide sequence that encodes the protein product of
choice.
51. The host cell of claim 50 wherein the exogenous polynucleotide
is integrated into the chromosome of the host cell.
52. The host cell of claim 50 wherein the exogenous polynucleotide
is present in the host cell as part of a replicable vector.
53. A method for producing a protein product of choice, the method
comprising: (a) providing the host cell of claim 50; and (b)
growing the host cell; thereby to produce a cell culture or
recombinant organism comprising an increased level of the protein
product of choice compared to the level of production of the
protein product of choice achieved by growing, under the same
conditions, the same host cell that has not been genetically
modified to cause over-expression of one or more helper
proteins.
54. The method of claim 53 wherein the step of growing the host
cell involves culturing the host cell in a culture medium.
55. The method of claim 53 further comprising the step of purifying
the thus expressed protein product of choice from the cultured host
cell, recombinant organism or culture medium.
56. Method of preparing a the host cell of claim 32, by
transformation of a host cell with a polynucleotide, wherein the
polynucleotide comprises a sequence encoding a helper protein
selected from the list comprising (a) a chaperone selected from a
DnaJ-like protein an Hsp70 family protein, and SIL1, and wherein
the DnaJ-like protein is not SCJ1; and (b) a protein involved in
the formation of disulphide bonds in other proteins selected from
ERO1, ERV2, EUG1, MPD1, MPD2, EPS1 and PDI1.
Description
FIELD OF THE INVENTION
[0001] The present application relates to gene expression
techniques.
BACKGROUND OF THE INVENTION
[0002] The listing or discussion of a prior-published document in
this specification should not necessarily be taken as an
acknowledgement that the document is part of the state of the art
or is common general knowledge.
[0003] A key parameter in the development of a commercially viable
process for the production of a recombinant protein is the yield of
the product from the host organism.
[0004] Factors that influence the yield of a particular
heterologous protein are complex and include the biochemical and
biophysical properties of the protein itself; its influence on, and
modification of, the host's own cellular functions; and the choice
and deployment of those sequences that are necessary for efficient
transcription, translation, secretion (if required) and plasmid
stability.
SUMMARY OF THE INVENTION
[0005] We have identified a series of proteins (hereinafter
"helper" proteins) that are over-expressed in a (non-publicly
available) S. cerevisiae that possesses increased production of a
protein product of choice, such as a recombinant protein. These
overexpressed helper proteins have all, individually, been
previously identified.
[0006] In the case of some of these helper proteins, there is
nothing in the art to suggest that their over-expression would aid
in the increased production of a recombinant heterologous protein
product of choice.
[0007] In the case of some of the other identified helper proteins,
the (as yet unpublished) art has recognised that their
over-expression can aid in increasing the production of a
recombinant heterologous protein product of choice (see
PCT/GB2004/005462). However, there is nothing in the art to suggest
that the combined and simultaneous over-expression of such helper
proteins would further enhance the production of a protein product
of choice.
[0008] Accordingly, the present invention provides a host cell
suitable for enhanced production of a protein product of choice
wherein the host cell is genetically modified to cause
over-expression of one or more of the identified helper
proteins.
[0009] Thus the present invention provides a host cell that is
suitable for enhanced production of a protein product of choice
characterised in that the host cell comprises a first gene encoding
a first helper protein as defined herein, or a variant thereof, and
a second gene encoding a desired protein product of choice, wherein
the host cell is genetically modified to cause over-expression of
the first helper protein, and-- [0010] (a) wherein the first and
second genes are not both present within the host cell on the same
2 .mu.m-family plasmid (and optionally the first gene is not
present within the host cell on any 2 .mu.m-family plasmid; and
further optionally the second gene is not present within the host
cell on any 2 .mu.m-family plasmid); and [0011] (b) wherein the
host cell is not genetically modified to cause over-expression of a
further helper protein that is different from the first helper
protein and is selected from the group consisting of AHA1, CCT2,
CCT3, CCT4, CCT5, CCT6, CCT7, CCT8, CNS1, CPR3, CPR6, ERO1, EUG1,
FMO1, HCH1, HSP10, HSP12, HSP104, HSP26, HSP30, HSP42, HSP60,
HSP78, HSP82, JEM1, MDJ1, MDJ2, MPD1, MPD2, PDI1, PFD1, ABC1, APJ1,
ATP11, ATP12, BTT1, CDC37, CPR7, HSC82, KAR2, LHS1, MGE1, MRS11,
NOB1, ECM10, SSA1, SSA2, SSA3, SSA4, SSC1, SSE2, SIL1, SLS1, ORM1,
ORM2, PER1, PTC2, PSE1, UBI4 and HAC1 or a truncated intronless
HAC1 (and optionally, the host cell is not genetically modified to
cause over-expression of any further helper protein that is
different from the first helper protein).
[0012] The thus over-expressed first helper protein may be any
helper protein defined below. For example, the over-expressed first
helper protein may be a DnaJ-like protein (such as JEM1), an Hsp70
family member protein (such as LHS1) or SIL1, or a variant of any
of these. Over-expression of the first helper protein may be
achieved by any suitable means of genetic modification known in the
art. Suitable examples of such approaches for genetic modification
are discussed in more detail below.
[0013] The host cell may or may not comprise a recombinant copy,
such as a plasmid encoded copy, or a chromosomally integrated
recombinant copy, of a gene encoding the further helper protein as
defined in (b) above. Thus, in one embodiment, the first helper
protein may be the only helper protein that is over-expressed by
the host cell.
[0014] In another embodiment, the invention provides a host cell
that is suitable, for enhanced production of a protein product of
choice characterised in that the host cell is genetically modified
to cause over-expression of a helper protein selected from the list
comprising SCJ1, FKB2, SSE1, ERV2, DER1, DER3, HRD3, UBC7 and DOA4.
The host cell may or may not be genetically modified to cause
over-expression of two or more helper proteins, at least one of
which is a helper protein selected from the list comprising SCJ1,
FKB2, SSE1, ERV2, DER1, DER3, HRD3, UBC7 and DOA4. In that case, at
least one other helper may or may not be selected from the list
comprising-- [0015] (a) chaperones selected from a DnaJ-like
protein (such as JEM1), an Hsp70 family member protein (such as
LHS1), SCJ1, KAR2, SIL1 (note that, SIL1 has previously been
referred to as SLS1), FKB2, SSA1, SSA2, SSA3, SSA4, SSE1, SSE2,
SSB1, SSB2, ECM10, MDJ1 and MDJ2. [0016] (b) proteins involved in
the formation of disulphide bonds in other proteins selected from
ERO1, ERV2, EUG1, MPD1, MPD2, EPS1 and PDI1; [0017] (c) proteins
involved in protein degradation selected from DER1, DER3, HRD3,
UBC7 and DOA4; and [0018] (d) HAC1.
[0019] For example, the host cell may or may not be genetically
modified to cause over-expression of two or more helper proteins
selected from a DnaJ-like protein (such as JEM1), an Hsp70 family
protein (such as LHS1) and SIL1. For example, the host cell
according to may or may not be genetically modified to cause
over-expression of-- [0020] (a) a DnaJ-like protein and an Hsp70
family protein; or [0021] (b) a DnaJ-like protein and SIL1; or
[0022] (c) an Hsp70 family protein and SIL1.
[0023] The host may or may not be genetically modified to cause
over-expression of three or more helper proteins, wherein the three
or more helper proteins comprise a DnaJ-like protein, an Hsp70
family protein and SIL1 for example JEM1, LHS1 and SIL1.
[0024] The Hsp70 family protein may or may not be a protein that
localises to the lumen of the ER. The Hsp70 family protein may or
may not be a prokaryotic Hsp70 family protein. The Hsp70 family
protein may or may not be a eukaryotic Hsp70 family protein. The
Hsp70 family protein may or may not be LHS1, KAR2, SSA1, SSA2,
SSA3, SSA4, SSE1, SSE2, SSB1, SSB2 or ECM10, such as from yeast,
for example, from S. cerevisiae. LHS1 may or may not be a preferred
Hsp70 family protein for use in the present invention. Other Hsp70
family proteins for use in the present invention may or may not
include a mammalian BiP (GRP78) (, such as the protein described by
Haas and Wabl (1983) Nature 306, 387), a mammalian HSP72 (HSP70),
HSP73 (HSC70) or mtp70, a mammalian GRP170 (such as the protein
described by Lin et al (1993) Mol. Biol. Cell 4, 1109), a mammalian
HSP70 protein (such as a protein as reviewed by Ohtsuka and Hata.
(2000) International Journal of Hyperthermia 16, 231; Gething and
Sambrook (1992) Nature 355, 33; and/or Craig and Gross (1991) TIBS
16, 135), a Gallus gallus HSP70 protein, such as the protein
defined by accession number to AAO44921 (Mazzi et al (2003) Genet.
Mol. Biol. 26, 275-281), a Nicotiana tabacum luminal binding
protein (BiP), such as the protein defined by accession number
CAA42661 (Denecke et al (1991) Plant Cell 3, 1025), a Paramecium
caudatum HSP70 protein, such as the protein defined by accession
number BAE16705 (Hori et al (2006) Mol. Phylogenet. Evol. 38, 697),
a Hordeum vulgare HSP70 protein, such as a subsp. vulgare HSP70
protein accession number, such as the protein defined by AAA62325
(Chen et al (1994) Plant Physiol. 106, 815), an Arabidopsis
thaliana HSP70 protein accession number NP.sub.--187864, the
Chlamydia trachomatis A/HAR-13 chaperone protein dnaK (Heat shock
protein 70) (Heat shock 70 kDa protein) (HSP70), such as the
protein defined by accession number Q3KLV7 (Carlson et al (2005)
Infect. Immun. 73, 6407), a Pongo pygmaeus hsp70 protein, such as
the protein defined by accession number CAH92327, a Haemophilus
influenzae 86-028NP HSP70 protein, such as the protein defined by
accession number YP.sub.--249343 (Harrison et al (2005) J.
Bacteriol. 187, 4627), a Streptococcus pneumoniae HSP70 protein,
such as the protein defined by accession number AAB39221, a Mus
musculus HSP70 protein, such as the protein defined by accession
number AAC84169 (Xie et al (2003) Genome Res. 13, 2621), a Bacillus
subtilis HSP70 protein, such as the protein defined by accession
number BAA12464 (Mizuno et al. (1996) Microbiology (Reading, Engl.)
142, 3103), and a Escherichia coli DnaK protein, such as the
protein defined by Slepenkov and Witt (2002) Mol. Microbiol. 45,
1197. It will be appreciated that, in the rest of this
specification, reference to LHS1 may or may not be taken to be, by
extension, a reference to an equivalent Hsp70 family protein, such
as an Hsp70 family protein as defined in this paragraph.
[0025] Other preferred Hsp70 family proteins may have an activity
equivalent to LHS1, when co-expressed with one or both of JEM1 and
SIL1 for example in the manner as set out in the present examples.
Thus, a host cell of the present invention, when genetically
modified to cause simultaneous over-expression of a preferred Hsp70
family protein with one or both of JEM1 and SIL1, will provide at
least substantially the same increase in the production of a
protein product and/or at least substantially the same reduction of
fragmentation of a protein product, as is observed in the same host
cell when genetically modified to cause simultaneous
over-expression of LHS1 with one or both of JEM1 and SIL1 the
increase being compared to the to the level of production of the
same protein product, and/or the level of fragmentation of the same
protein product, in the same host cell that has not been
genetically modified to cause overexpression of any of LHS1, JEM1
or SIL1.
[0026] By "substantially the same increase in the production of a
protein product", we, mean at least 10%, 20%, 30%, 40%, 50%, 60%,
70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, substantially 100% or
greater than 100% of the increase in production of a protein
product that is observed when the host cell is genetically modified
to cause simultaneous over-expression of LHS1 with one or both of
JEM1 and SIL1 (the increased being compared to the level of
production of the same protein product in the same host cell that
has not been genetically modified to cause overexpression of any of
LHS1, JEM1 or SIL1).
[0027] By "substantially the same reduction of fragmentation of a
protein product", we mean at least 10%, 20%, 30%, 40%, 50%, 60%,
70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, substantially 100% or
greater than 100% of the reduction of fragmentation of a protein
product that is observed when the host cell is genetically modified
to cause simultaneous over-expression of LHS1 with one or both of
JEM1 and SIL1 (the reduction of fragmentation of a protein product
being compared to the level of fragmentation of the same protein
product in the same host cell that has not been genetically
modified to cause overexpression of any of LHS1, JEM1 or SIL1).
[0028] DnaJ-like proteins are reviewed in Walsh et al, 2004, EMBO
reports, 5, 567-571. The DnaJ-like protein typically comprises a
J-domain as defined in Walsh et al, 2004, op. cit. the contents of
which are incorporated herein by reference. The DnaJ-like protein
may or may not be a prokaryotic DnaJ-like protein. The DnaJ-like
protein may or may not be a eukaryotic DnaJ-like protein. The
DnaJ-like protein may or may not be any one of the yeast DnaJ
proteins such as a protein selected from JEM1, MDJ1, MDJ2, SEC63,
YDJ1, XDJ1, APJ1, SIS1, DJP1, ZUO1, SWA2, JJJ1, JJJ2, JJJ3, CAJ1,
CWC23, PAM18, JAC1, JID1, SCJ1, HLJ1 to and ERJ5. The DnaJ-like
protein may or may not be a protein that localises to the ER, such
as JEM1, SCJ1, HLJ1, SEC63 or ERJ5, and may or may not be a protein
that localises to the ER membrane. The DnaJ-like protein may or may
not be a protein that localises to the cytoplasm of the host cell,
such as YDJ1, XDJ1, APJ1, SIS1, DJP1, ZUO1, SWA2, JJJ1, JJJ2 or
JJJ3. The DnaJ-like protein may or may not be a protein that
localises to the nucleoplasm of the host cell, such as CAJ1 or
CWC23. The DnaJ-like protein may or may not be a protein that
localises to the mitochondria of the host cell, such as MDJ1, MDJ2,
PAM18, JAC1 or JID1. The DnaJ-like protein is typically not SCJ1.
JEM1 may or may not be a preferred DnaJ-like protein for use in the
present invention. Other DnaJ-like proteins may or may not include
the following proteins or proteins families, or fragments or
variants thereof-- [0029] the HSP40 class of proteins (reviewed by
Ohtsuka and Hata. (2000) International Journal of Hyperthermia 16,
231 and Table 1 therein); [0030] a mammalian Erdj1 (such as MTJ1,
Chevalier et al (2000) J. Biol. Chem. 275 19620); [0031] a
mammalian Erdj2 such as hSec63, Skowronek et al (1999) J. Biol.
Chem. 380, 1133); [0032] a mammalian Erdj3 (such as HEDJ/Scj1p,
Shen and Hendershot (2005) Mol. Biol. Cell. 16, 40); [0033] a
mammalian Erdj4 (such as described in Shen et al (2002) J. Biol.
Chem. 277, 15947); [0034] a mammalian Erdj5 (such as described in
Cunnea et al (2003) J. Biol. Chem. 278, 1059); [0035] a Gallus
gallus DnaJ homolog subfamily B member 11 precursor, such as the
ER-associated dnaJ protein 3 ErJ3, the ER-associated Hsp40
co-chaperone (hDj9, or the PWP1-interacting protein 4, such as
defined by accession number XP.sub.--422682; [0036] a Nicotiana
tabacum DnaJ homolog, such as the protein defined by accession
number BAC53943; [0037] a Arabidopsis thaliana DnaJ homolog, such
as the protein defined by accession number AAB49030 (Zhou et al
(1999) Plant Physiol. 121, 1053); [0038] a Chlamydia trachomatis
A/HAR-13 Chaperone protein dnaJ, such as the protein defined by
accession number YP.sub.--328153 (Carlson et al (2005) Infect.
Immun. 73, 6407); [0039] a Pongo pygmaeus DnaJ homolog subfamily B
member 9, such as the protein defined by accession number Q5R9A4;
[0040] a Haemophilus influenzae Rd KW20 Dna-J like membrane
chaperone protein, such as the protein defined by accession number
NP.sub.--438440 (Fleischmann et al (1995) Science 269, 496); [0041]
a Escherichia coli DnaJ protein, such as the protein defined by
accession number AAA00009 (Ohki et al (1986) J. Biol. Chem. 261,
1778); [0042] a Escherichia coli DnaJ-like protein, such as the
protein defined by accession number BAB96590 (Musso et al (1977)
Proc. Natl. Acad. Sci. U.S.A. 74, 106); [0043] a Streptococcus
pneumoniae DnaJ protein, such as the protein defined by accession
number AAB39222; [0044] a Mus musculus DnaJ homolog, such as a
subfamily B member 6 (Heat shock protein. J2) (HSJ-2) (MRJ) (mDj4)
such as the protein defined by accession number XP.sub.--987742;
[0045] a Bacillus subtilis DnaJ protein, such as the protein
defined by accession number BAA12465 (Mizuno et al (1996)
Microbiology (Reading Engl.) 142, 3103); and [0046] a plant Sorghum
bicolour DNAJ domain protein, such as the protein defined by
accession number ABF48023.
[0047] It will be appreciated that, in the rest of this
specification, reference to JEM1 may or may not be taken to be, by
extension, a reference to an equivalent DnaJ-like protein, such as
a DnaJ-like protein as defined in the above paragraph.
[0048] Other preferred DnaJ-like proteins may have an activity
equivalent to JEM1, when co-expressed with one or both of LHS1 and
SIL1, for example in the manner as set out in the present examples.
Thus, a host cell of the present invention, when genetically
modified to cause simultaneous over-expression of a preferred
DnaJ-like protein with one or both of LHS1 and SIL1, will provide
at least substantially the same increase in the production of a
protein product and/or at least substantially the same reduction of
fragmentation of a protein product, as is observed in the same host
cell when genetically modified to cause simultaneous
over-expression of JEM1 with one or both of LHS1 and SIL1, the
increase being compared to the level of production of the same
protein product, and/or the level of fragmentation of the same
protein product, in the same host cell that has not been
genetically modified to cause overexpression of any of LHS1, JEM1
or SIL1.
[0049] By "substantially the same increase in the production of a
protein product", we mean at least 10%, 20%, 30%, 40%, 50%, 60%,
70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, substantially 100% or
greater than 100% of the increase in production of a protein
product that is observed when the host cell is genetically modified
to cause simultaneous over-expression of JEM1 with one or both of
LHS1 and SIL1 (the increase being compared to the level of
production of the same protein product in the same host cell that
has not been genetically modified to cause overexpression of any of
LHS1, JEM1 or SIL1).
[0050] By "substantially the same reduction of fragmentation of a
protein product", we mean at least 10%, 20%, 30%, 40%, 50%, 60%,
70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, substantially 100% or
greater than 100% of the reduction of fragmentation of a protein
product that is observed when the host cell is genetically modified
to cause simultaneous over-expression of JEM1 with one or both of
LHS1 and SIL1 (the reduction of fragmentation of a protein product
being compared to the level of fragmentation of the same protein
product in the same host cell that has not been genetically
modified to cause overexpression of any of LHS1, JEM1 or SIL1).
[0051] The host cell that is genetically modified to cause
over-expression of two or more; such as at least three, helper
proteins selected from a DnaJ-like protein, an Hsp70 family protein
and SIL1 may or may not be further genetically modified to cause
over-expression of at least one, two, three, four, five, six or
seven proteins involved in the formation of disulphide bonds in
other proteins selected from the group consisting of ERO1, ERV2,
EUG1, MPD1, MPD2, EPS1 and PDI1. PDI1 may or may not be
preferred.
[0052] In another embodiment, the invention provides a host cell
suitable for enhanced production of a protein product of choice
characterised in that the host cell is genetically modified to
cause over-expression of three or more helper proteins, wherein the
three or more helper proteins are selected from the list
comprising-- [0053] (a) chaperones selected from a DnaJ-like
protein (such as JEM1), an Hsp70 family member protein (such as
LHS1), SCJ1, KAR2, SIL1, FKB2, SSA1, SSA2, SSA3, SSA4, SSE1, SSE2,
SSB1, SSB2, ECM10, MDJ1 and MDJ2. [0054] (b) proteins involved in
the formation of disulphide bonds in other proteins selected from
ERO1, ERV2, EUG1, MPD1., MPD2, EPS1 and PDI1; [0055] (c) proteins
involved in protein degradation selected from DER1, DER3, HRD3,
UBC7 and DOA4; and [0056] (d) HAC1.
[0057] The three or more helper proteins may or may not comprise at
least one, two, three, four, five, six, seven, eight, nine, ten,
eleven, twelve, thirteen, fourteen, fifteen, sixteen or seventeen
of the chaperones selected from the group consisting of JEM1, an
Hsp70 family member protein (such as LHS1), SCJ1, KAR2, SIL1, FKB2,
SSA1, SSA2, SSA3, SSA4, SSE1, SSE2, SSB1, SSB2, ECM10, MDJ1 and
MDJ2. The three or more helper proteins may or may not comprise at
least one, two, three, four, five, six or seven proteins involved
in the formation of disulphide bonds in other proteins selected
from the group consisting of ERO1, ERV2, EUG1, MPD1, MPD2, EPS1 and
PDI1. The three or more helper proteins may or may not comprise at
least one, two, three, four or five of the proteins involved in
protein degradation selected from DER1, DER3, HRD3, UBC7 and
DOA4.
[0058] It will be appreciated that the host cell may or may not
comprise a polynucleotide sequence that encodes a protein product
of choice.
[0059] In one embodiment, the host cell comprises a polynucleotide
sequence that encodes a protein product of choice. The protein
product of choice may or may not be a protein that is naturally
produced by the host cell or may or may not be a heterologous
protein. In this context, a "heterologous protein" is a protein
that is not naturally encoded by the host cell. The polynucleotide
sequence that encodes the protein product of choice may or may not
be an endogenous polynucleotide sequence or (in particular, where
the protein product of choice is a heterologous protein) the
polynucleotide sequence that encodes the protein product of choice
may or may not be an exogenous polynucleotide, and the exogenous
polynucleotide may or may not be integrated into the chromosome of
the host cell or present in the host cell as part of a replicable
vector, such as a plasmid.
[0060] However, the present invention also contemplates the
production of host cells suitable for enhanced production of a
protein product of choice, into which an appropriate polynucleotide
sequence, encoding the protein product of choice, can be later
introduced. Therefore, in another embodiment, the host cell does
not comprise a polynucleotide sequence that encodes a protein
product of choice.
[0061] Suitable host cells are discussed below.
[0062] By "enhanced production" we include the meaning that the
level of production of protein product of choice is greater in a
cultured population of the genetically modified host cell than in a
cultured population of the same host cell that has not been
genetically modified to cause over-expression of one or more of the
identified helper proteins. Typically, the measurement can be made
under culture conditions that are standard for the growth of the
host cell that is being used.
[0063] Thus the production of the protein product of choice in a
cultured population of the genetically modified host cell of the
invention be greater than, typically at least 1%, 2%, 3%, 4%, 5%,
6%, 7%, 8%, 9%, 10% (i.e. 1.1-fold), 20% (i.e. 1.2-fold), 30% (i.e.
1.3-fold), 40% (i.e. 1.4-fold), 50% (i.e. 1.5-fold), 60% (i.e.
1.6-fold), 70% (i.e. 1.7 fold), 80% (i.e. 1.8-fold), 90% (i.e.
1.9-fold), 100% (i.e. 2-fold), 3-fold, 4-fold, 5-fold, 6-fold,
7-fold, 8-fold, 9-fold, 10-fold, 20-fold, 30-fold, 40-fold,
50-fold, 60-fold, 70-fold, 80-fold, 90-fold, 100-fold, 200-fold,
300-fold, 400-fold, 500-fold, 600-fold, 700-fold, 800-fold,
900-fold, or 1000-fold greater than, the production of the protein
product of choice in a cultured population of the same host cell
that has not been genetically modified to cause over-expression of
one or more of the identified helper proteins. These figures may,
or may not, be figures that have been normalised to account for
differences in the cell growth of the two cultured populations, as
compared.
[0064] For example, the production of the protein product of choice
in a cultured population of the genetically modified host cell of
the invention may be up to 10% (i.e. 1.1-fold), 20% (i.e.
1.2-fold), 30% (i.e. 1.3-fold), 40% (i.e. 1.4-fold), 50% (i.e.
1.5-fold), 60% (i.e. 1.6-fold), 70% (i.e. 1.7 fold), 80% (i.e.
1.8-fold), 90% (i.e. 1.9-fold), 100% (i.e. 2-fold), 3-fold, 4-fold,
5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold,
30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold,
100-fold, 200-fold, 300-fold, 400-fold, 500-fold, 600-fold,
700-fold, 800-fold, 900-fold, 1000-fold or 2000-fold greater than
the production of the protein product of choice in a cultured
population of the same host cell that has not been genetically
modified to cause over-expression of one or more of the identified
helper proteins. These figures may, or may not, be figures that
have been normalised to account for differences in the cell growth
of the two cultured populations, as compared.
[0065] Typically, the protein product of choice may be produced in
a cultured population of the genetically modified host cell of the
invention to produce a culture containing at least 0.001 gL.sup.-1,
such as at least 0.01 at least 0.1 gL.sup.-1, 1 gL.sup.-1, 2
gL.sup.-1, 3 gL.sup.-1, 4 gL.sup.-1, 5 gL.sup.-1, 6 gL.sup.-1, 7
gL.sup.-1, 8 gL.sup.-1, 9 gL.sup.-1, 10 gL.sup.-1, 20 gL.sup.-1, 30
gL.sup.-1, 40 gL.sup.-1, 50 gL.sup.-1, 60 gL.sup.-1, 70 gL.sup.-1,
80 gL.sup.-1, 90 gL.sup.-1, or 100 gL.sup.-1 of the protein product
of choice. The protein product of choice may be produced in a
cultured population of the genetically modified host cell of the
invention to produce a culture containing up to 0.01 gL.sup.-1, 0.1
gL.sup.-1, 2 gL.sup.-1, 3 gL.sup.-1, 4 gL.sup.-1, 5 gL.sup.-1, 6
gL.sup.-1, 7 gL.sup.-1, 8 gL.sup.-1, 9 gL.sup.-1, 10 gL.sup.-1, 20
gL.sup.-1, 30 gL.sup.-1, 40 gL.sup.-1, 50 gL.sup.-1, 60 gL.sup.-1,
70 gL.sup.-1, 80 gL.sup.-1, 90 gL.sup.-1, 100 gL.sup.-1 or 200
gL.sup.-1 of the protein product of choice.
[0066] By "enhanced production" we also include the meaning that
the level of activity of the protein product of choice that is
produced by the host cell is greater in a cultured population of
the genetically modified host cell than in a cultured population of
the same host cell that has not been genetically modified to cause
over-expression of one or more of the identified helper proteins.
The nature of the activity will depend on the identity of the
protein product of choice and may, for example, be a measurement of
the catalytic activity of the protein upon a substrate or the
binding properties of the protein to a ligand. Typically, the
measurement of protein activity can be made under culture
conditions that are standard for the growth of the host cell that
is being used or following isolation of the protein from the
culture medium. In either case, the comparison should be made on
the basis of activity per unit volume of culture or protein
recovered therefrom. The comparison may, or may not, be normalised
to account for differences in the cell growth of the two cultured
populations, as compared.
[0067] Thus the activity of the protein product of choice that is
produced in a cultured population of the genetically modified host
cell of the invention may be greater than, typically at least 10%
(i.e. 1.1-fold), 20% (i.e. 1.2-fold), 30% (i.e. 1.3-fold), 40%
(i.e. 1.4-fold), 50% (i.e. 1.5-fold), 60% (i.e. 1.6-fold), 70%
(i.e. 1.7-fold), 80% (i.e. 1.8-fold), 90% (i.e. 1.9-fold), 100%
(i.e. 2-fold), 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold,
9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold, 60-fold,
70-fold, 80-fold, 90-fold, 100-fold, 200-fold, 300-fold, 400-fold,
500-fold, 600-fold, 700-fold, 800-fold, 900-fold, 1000-fold,
10.sup.4-fold, 10.sup.5-fold, or 10.sup.6-fold greater than, the
activity of the protein product of choice in a cultured population
of the same host cell that has not been genetically modified to
cause over-expression of one or more of the identified helper
proteins.
[0068] For example, the activity of the protein product of choice
in a cultured population of the genetically modified host cell of
the invention may be up to 10% (i.e. 1.1-fold), 20% (i.e.
1.2-fold), 30% (i.e. 1.3-fold), 40% (i.e. 1.4-fold), 50% (i.e.
1.5-fold), 60% (i.e. 1.6-fold), 70% (i.e. 1.7 fold), 80% (i.e.
1.8-fold), 90% (i.e. 1.9-fold), 100% (i.e. 2-fold), 3-fold, 4-fold,
5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold,
30-fold, 40-fold, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold,
100-fold, 200-fold, 300-fold, 400-fold, 500-fold, 600-fold,
700-fold, 800-fold, 900-fold, 1000-fold, 10.sup.4-fold,
10.sup.5-fold, or 10.sup.6-fold greater than the activity of the
protein product of choice in a cultured population of the same host
cell that has not been genetically modified to cause
over-expression of one or more of the identified helper
proteins.
[0069] By "enhanced production" we include the additional or
alternative meaning that the level of degradation of the protein
product of choice is reduced when produced by a cultured population
of the genetically modified host cell of the present invention
compared to the level of degradation of the protein product of
choice when produced by a cultured population of the same host cell
that has not been genetically modified to cause over-expression of
one or more of the identified helper proteins according to the
present invention. The level of protein degradation can be
determined by quantification of fragments of the protein product of
choice relative to the total of the protein product of choice, for
example when by analysis of SDS-PAGE using densitometry. When
expressed as a percentage of detected protein product fragments
relative to total protein product levels detected (i.e. total
protein product detected=full length protein product+degradation
products) then the percentage of detected protein product fragments
when produced by a cultured population of the genetically modified
host cell of the present invention may be, or be less than, 99%,
98%, 97%, 96%, 05%, 04%, 03%, 92%, 90%, 85%, 80%, 75%, 70%, 65%,
60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%,
6%, 5%, 4%, 3%, 2%, 1% or less, such as up to 98%, 97%, 96%, 95%,
94%, 93%, 92%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%,
40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%,
1% or less of the percentage of detected protein product fragments
when produced by a cultured population of the same host cell that
has not been genetically modified to cause over-expression of one
or more of the identified helper proteins according to the present
invention. These values may or may not be normalised, for example
based on culture optical density readings, to account for different
growth rates observed between strains.
[0070] By "enhanced production" we include the additional or
alternative meaning that the level of post-translational
modification of the protein product of choice is increased or
reduced when produced by a cultured population of the genetically
modified host cell of the present invention compared to the level
of post-translational modification of the protein product of choice
when produced by a cultured population of the same host cell that
has not been genetically modified to cause over-expression of one
or more of the identified helper proteins according to the present
invention. For example, the altered (i.e. increased or reduced)
level of post-translational modification may be an alteration in
the level of proteolytic cleavage, hexosylation (for example
mannosylation), glycosylation, phosphorylation,
phosphopantetheinylation, carbamylation, carboxylation (such as
.gamma.-carboxylation), sialation, sulphonation, hydroxylation,
prenylation, isoprenylation, acylation, ubiquitination,
lipoylation, biotinylation, glycylation, glutamylation,
methylation, alkylation, acetylation, formylation, selenation,
disulphide bond formation or oligomerisation of the protein product
of choice. The level of post-translational modification of the
protein product of choice can be determined by methods well known
in the art, such as by mass spectrometry techniques (for example,
see Larsen et al, 2006, BioTechniques, 40, 790-798) well known in
the art.
[0071] By "enhanced production" we include the additional or
alternative meaning that the level of stress experienced by a cell
that is being cultured to produce the protein product of choice is
reduced, compared to the level of stress experienced by a cultured
population of the same host cell that has not been genetically
modified to cause over-expression of one or more of the identified
helper proteins according to the present invention. For example,
"enhanced production" can include the additional or alternative
meaning that the unfolded protein response is reduced in a host
cell. The level of stress, and the level of the unfolded protein
response, can be measured by determination of the proportion of
HAC1.sup.i to total HAC1 transcript levels. Total HAC1 transcript
levels are the sum of HAC1.sup.i transcript levels and unspliced
HAC1 (HAC1.sup.u) transcript levels in a cell. A reduced proportion
of HAC1.sup.i transcript levels compared to total HAC1 transcript
level, relative to a control, is indicative of reduced stress and
reduced. UPR signalling relative to that control. Helper proteins
suitable for achieving this effect may include Hsp70 family
proteins (such as LHS1) and DnaJ-like proteins (such as JEM1) and
combinations of other helper proteins such as disclosed in the
present application.
[0072] In principle, any "protein product of choice" can be
produced. The identity of preferred embodiments of the "protein
product of choice" is discussed further below.
[0073] The host cell is genetically modified to cause
over-expression of one or more of the helper proteins. By
"over-expression", in the context of helper proteins, we mean that
the measurable level of mRNA encoding the one or more helper
proteins, and/or the measurable level of the one or more helper
proteins themselves, and/or the measurable level of the helper
protein activity, is greater than the measurable level in a host
cell that has not been genetically modified. Typically, the
measurement will be made under culture conditions that are standard
for the growth of the host cell that is being used. Standard
conditions for yeast cell growth are discussed, for example, in WO
96/37515, WO 00/44772 and WO 99/00504, the contents of which are
incorporated herein by reference.
[0074] Thus the host cell may or may not be genetically modified to
cause a level of expression of one or more of the helper proteins
that is at least a 1.1-fold, 1.2-fold, 1.3-fold, 1.4-fold,
1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold,
9-fold, 10-fold, 20-fold, 30-fold, 40-fold, 50-fold or more, of
unmodified level of expression of one or more of the helper
proteins.
[0075] For example, the host cell may or may not be genetically
modified to cause a level of expression of one or more of the
helper proteins that is up to 1.2-fold, 1.3-fold, 1.4-fold,
1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold,
9-fold, 10-fold, 15-fold, 20-fold, 30-fold, 40-fold, 50-fold,
60-fold, 70-fold, 80-fold, 90-fold or 100-fold of the
unmodified-type level of expression of one or more of the helper
proteins.
[0076] For example, the host cell may be genetically modified to
cause a level of expression of one or more of the helper proteins
that is between 1- to 30-fold, such as about 2- to 25-fold, of the
unmodified-type level of expression of one or more of the helper
proteins.
[0077] The host cell may or may not be genetically modified to
cause over-expression of one or more of the helper proteins by the
introduction of one or more recombinant copies of one or more
polynucleotides that each comprise a region (the "coding region",
or "open reading frame", which can be abbreviated to "ORF") that
encodes one or more helper proteins.
[0078] A copy of the polynucleotide may or may not be introduced
into the chromosome of the host cell and/or may or may not be
encoded by a plasmid or other vector that is used to transform the
host cell.
[0079] The polynucleotide may or may not comprise some or all of
the regulatory sequences necessary to cause transcription and/or
translation of the ORF of the polynucleotide.
[0080] Regulatory sequences necessary to cause transcription and/or
translation of the ORF of the polynucleotide include sequences that
modulate (i.e., promotes or reduces, typically promotes) the
expression (i.e., the transcription and/or translation) of an ORF
to which it is operably linked. Regulatory regions typically
include promoters, terminators, ribosome binding sites and the
like. The skilled person will appreciate that the choice of
regulatory region will depend upon the intended expression system.
For example, promoters may or may not be constitutive or inducible
and may or may not be cell- or tissue-type specific or
non-specific.
[0081] Suitable regulatory regions, may be about, or up to, 5 bp,
10 bp, 15 bp, 20 bp, 25 bp, 30 bp, 35 bp, 40 bp, 45 bp, 50 bp, 60
bp, 70 bp, 80 bp, 90 bp, 100 bp, 120 bp, 140 bp, 160 bp, 180 bp,
200 bp, 220 bp, 240 bp, 260 bp, 280 bp, 300 bp, 350 bp, 400 bp, 450
bp, 500 bp, 550 bp, 600 bp, 650 bp, 700 bp, 750 bp, 800 bp, 850 bp,
900 bp, 950 bp, 1000 bp, 1100 bp, 1200 bp, 1300 bp, 1400 bp, 1500
bp or greater, in length.
[0082] Such non-coding regions and regulatory regions are not
restricted to the native non-coding regions and/or regulatory
regions naturally associated with the ORF.
[0083] Where the host cell is yeast, such as Saccharomyces
cerevisiae, suitable promoters for S. cerevisiae include those
associated with the PGK1 gene, GAL1 or GAL10 genes, TEF1, TEF2,
PYK1, PMA1, CYC1, PHO5, TRP1, ADH1; ADH2, the genes for
glyceraldehyde-3-phosphate dehydrogenase (for example, TDH1, TDH2
or TDH3), hexokinase (for example, HXK1 or HXK2), pyruvate
decarboxylase (for example, PDC1, PDC5 or PDC6),
phosphofructokinase (for example, PFK1 or PFK2), triose phosphate
isomerase (for example, TPI1), phosphoglucose isomerase (for
example, PGI1), glucokinase (for example, GLK1), .alpha.-mating
factor pheromone (for example, MF.alpha.-1 or MF.alpha.-2),
.alpha.-mating factor pheromone (for example, MFA1 or MFA2), PRB1,
PRA1, GPD1, and hybrid promoters involving hybrids of parts of 5'
regulatory regions with parts of 5' regulatory regions of other
promoters or with upstream activation sites (e.g. the promoter of
EP-A-258 067). Where multiple ORFs are to be expressed, a different
promoter may or may not be chosen for each ORF. The skilled person
can readily determine appropriate combinations of promoters. For
example, the promoters from the ADH1, PGK1, TDH1 and TEF1 genes are
used in combination to recombinantly over-express four helper
proteins in Example 3 below.
[0084] Suitable transcription termination signals are well known in
the art. Where the host cell is eukaryotic, the transcription
termination signal is preferably derived from the 3' flanking
sequence of a eukaryotic gene, which contains proper signals for
transcription termination and polyadenylation. Suitable 3' flanking
sequences may, for example, be those of the gene naturally linked
to the expression control sequence used, i.e. may correspond to the
promoter. Alternatively, they may be different. In that case, and
where the host is a yeast, preferably S. cerevisiae, then the
termination signal of the S. cerevisiae ADH1, ADH2, CYC1, or PGK1
genes are preferred.
[0085] It may be beneficial for the promoter and open reading frame
to be flanked by transcription termination sequences so that the
transcription termination sequences are located both upstream and
downstream of the promoter and open reading frame, in order to
prevent transcriptional read-through into neighbouring genes, and
visa versa.
[0086] In one embodiment, a suitable regulatory sequences in yeast,
such as Saccharomyces cerevisiae, includes: a yeast promoter (e.g.
the Saccharomyces cerevisiae PRB1 promoter), as taught in EP 431
880; and a transcription terminator, preferably the terminator from
Saccharomyces ADH1, as taught in EP 60 057. Other suitable
regulatory sequences are given in the examples, and include TEF1,
PGK1 and TDH1 promoters.
[0087] It may be beneficial for the non-coding region to
incorporate more than one DNA sequence encoding a translational
stop codon, such as UAA, UAG or UGA, in order to minimise
translational read-through and thus avoid the production of
elongated, non-natural fusion proteins. The translation stop codon
UAA is preferred. Preferably, the polynucleotide incorporates at
least two translation stop codons.
[0088] The term "operably linked" includes within its meaning that
a regulatory sequence is positioned within any non-coding region in
a gene such that it forms a relationship with an ORF that permits
the regulatory region to exert an effect on the ORF in its intended
manner. Thus a regulatory region "operably linked" to an ORF is
positioned in such a way that the regulatory region is able to
influence transcription and/or translation of the ORF in the
intended manner, under conditions compatible with the regulatory
sequence.
[0089] Alternatively, the polynucleotide may or may not be formed
in such a manner that it can take advantage of endogenous
regulatory sequences within the chromosome or plasmid to cause
transcription and/or translation of the coding region of the
polynucleotide. For example, the use of promoterless constructs is
well known in the art as a way of allowing an endogenous promoter
sequence to drive the expression of a recombinantly-introduced
polynucleotide coding region.
[0090] The skilled person will appreciate that the host cell may or
may not comprise endogenous copies of genes encoding one or more of
the helper proteins. Therefore, this invention also contemplates
genetic modifications to the host cell that cause increased steady
state levels of mRNA molecules encoding one or more helper proteins
and/or increased steady state levels of one or more helper
proteins.
[0091] This can include the genetic modification of operably linked
endogenous regulatory regions. For example, the endogenous promoter
in the gene of an endogenously encoded helper protein can be
replaced by a promoter that causes greater levels of expression of
the helper protein under culture conditions.
[0092] Alternatively, genetic modifications can be made to cis or
trans regulators of the gene of an endogenously encoded helper
protein, so as to increase the expression of the helper protein
under culture conditions. Thus, the polynucleotide region that
encodes a genetically encoded repressor of a gene of an
endogenously encoded helper protein could be genetically modified
to reduce or prevent repression of the endogenous helper protein
gene.
[0093] Alternative genetic modifications to increase the expression
of a helper protein or protein product of choice can involve
transient expression techniques known in to the art. For example,
suitable techniques are disclosed in Chen et al, 1997, Nucleic
Acids Research, 25, 4416-4418 and in Behr et al, 1989, Proc. Natl.
Acad. Sci. USA, 86, 6982-6986.
[0094] Thus, a number of techniques are available to the skilled
person to genetically modify a cell to cause over-expression of a
helper protein (and the same techniques may be used to cause
expression of a protein product of choice). Suitable techniques
include-- [0095] (i) introduction of a recombinant copy of an
encoding polynucleotide by integration into the chromosome of the
host cell (for example, either with associated regulatory sequences
or without associated regulatory sequences so as to take advantage
of endogenous regulatory sequences at the site of integration);
[0096] (ii) introduction of plasmid or other vector comprising a
recombinant copy of an encoding polynucleotide into the cell;
[0097] (iii) genetic modifications of a host cell's endogenous
regulatory region operably linked to the host cell's endogenous
copy of an ORF encoding a helper protein or protein product of
choice, to cause increased steady state levels of mRNA molecules
encoded by said ORF; [0098] (iv) genetic modifications to a cis or
trans regulator of the gene of an endogenously encoded helper
protein or protein product of choice; or [0099] (v) transient
expression of a helper protein or protein product of choice.
[0100] Where the host cell comprises a first gene encoding a
protein product of choice, and a second gene encoding a first
helper protein, then for example, [0101] the first gene may be a
gene as defined in (i) above and the second gene may be a gene as
defined in (i), (ii), (iii), (iv) or (v) above; [0102] the first
gene may be a gene as defined in (ii) above and the second gene may
be a gene as defined in (i), (ii), (iv) or (v) above (and where
both the first and second genes are introduced on plasmid or
vector, the first gene may or may not be introduced on the same
plasmid or vector as the to second gene); [0103] the first gene may
be a gene as defined in (iii) above and the second gene may be a
gene as defined in (i), (iv) or (v) above; [0104] the first gene
may be a gene as defined in (iv) above and the second gene may be a
gene as defined in (i), (iv) or (v) above; or [0105] the first gene
may be a gene as defined in (v) above and the second gene may be a
gene as defined in (i), (iv) or (v) above.
[0106] Where the host cell comprises a first gene encoding a
protein product of choice, and a second gene encoding a first
helper protein and a third gene encoding a second helper protein,
then for example, [0107] the first gene may be a gene as defined in
(i) above, and the second gene may be a gene as defined in (i)
above, and the third gene may be a gene as defined in (i), (iv) or
(v) above; [0108] the first gene may be a gene as defined in (i)
above, and the second gene may be a gene as defined in (ii) above,
and the third gene may be a gene as defined in (i), (ii), (iv) or
(v) above (and where both the second and third genes are introduced
on plasmid or vector, the second gene may or may not be introduced
on the same plasmid or vector as the third gene); [0109] the first
gene may be a gene as defined in (i) above, and the second gene may
be a gene as defined in (iii) above, and the third gene may be a
gene as defined in (i), (ii), (iv) or (v) above; [0110] the first
gene may be a gene as defined in (i) above, and the second gene may
be a gene as defined in (iv) above, and the third gene may be a
gene as defined in (i), (ii), (iii), (iv) or (v) above; [0111] the
first gene may be a gene as defined in (i) above, and the second
gene may be a gene as defined in (v) above, and the third gene may
be a gene as defined in (i), (ii), (iii), (iv) or (v) above; [0112]
the first gene may be a gene as defined in (ii) above, and the
second gene may be a gene as defined in (i) above, and the third
gene may be a gene as defined in (i), (iii), (iv) or (v) above (and
where both the first and third genes are introduced on plasmid or
vector, the first gene may or may not be introduced on the same
plasmid or vector as the third gene); [0113] the first gene may be
a gene as defined in (ii) above, and the second gene may be a gene
as defined in (ii) above, and the third gene may be a gene as
defined in (i), (iv) or (v) above (and where the first, second and
third genes are introduced on plasmid or vector, the first gene may
or may not be introduced on the same plasmid or vector as the
second gene, the first gene may or may not be introduced on the
same plasmid or vector as the third gene and the second gene may or
may not be introduced on the same plasmid or vector as the third
gene); [0114] the first gene may be a gene as defined in (ii)
above, and the second gene may be a gene as defined in (iii) above,
and the third gene may be a gene as defined in (i), (iii), (iv) or
(v) above; [0115] the first gene may be a gene as defined in (ii)
above, and the second gene may be a gene as defined in (iv) above,
and the third gene may be a gene as defined in (i), (iv) or (v)
above; [0116] the first gene may be a gene as defined in (ii)
above, and the second gene may be a gene as defined in (V) above,
and the third gene may be a gene as defined in (i), (iv) or (v)
above; [0117] the first gene may be a gene as defined in (iii)
above, and the second gene may be a gene as defined in (i) above,-
and the third gene may be a gene as defined in (i), (iii), (iv) or
(v) above; [0118] the first gene may be a gene as defined in (iii)
above, and the second gene may be a gene as defined in (ii) above,
and the third gene may be a gene as defined in (i), (iv) or (v)
above (and where both the second and third genes are introduced on
plasmid or vector, the second gene may or may not be introduced on
the same plasmid or vector as the third gene); [0119] the first
gene may be a gene as defined in (i) above, and the second gene may
be a gene as defined in (iii) above, and the third gene may be a
gene as defined in (i), (iv) or (v) above; [0120] the first gene
may be a gene as defined in (iii) above, and the second gene may be
a gene as defined in (iv) above, and the third gene may be a gene
as defined in (i), (iv) or (v) above; [0121] the first gene may be
a gene as defined in (iii) above, and the second gene may be a gene
as defined in (v) above, and the third gene may be a gene as
defined in (i), (iv) or (v) above; [0122] the first gene may be a
gene as defined in (iv) above, and the second gene may be a gene as
defined in (i) above, and the third gene may be a gene as defined
in (i), (ii), (iv) or (v) above; [0123] the first gene may be a
gene as defined in (iv) above, and the second gene may be a gene as
defined in (ii) above, and the third gene may be a gene as defined
in (i), (iii), (iv) or (v) above (and where both the second and
third genes are introduced on plasmid or vector, the second gene
may or may not be introduced on the same plasmid or vector as the
third gene); [0124] the first gene may be a gene as defined in (iv)
above, and the second gene may be a gene as defined in (iii) above,
and the third gene may be a gene as defined in (i), (iv) or (v)
above; [0125] the first gene may be a gene as defined in
(iv).above, and the second gene may be a gene as defined in (iv)
above, and the third gene may be a gene as defined in (i), (iv) or
(v) above; [0126] the first gene may be a gene as defined in (iv)
above, and the second gene may be a gene as defined in (v) above,
and the third gene may be a gene as defined in (i), (iv) or (v)
above; [0127] the first gene may be a gene as defined in (v) above,
and the second gene may be a gene as defined in (i) above, and the
third gene may be a gene as defined in (i), (ii), (iv) or (v)
above; [0128] the first gene may be a gene as defined in (v) above,
and the second gene may be a gene as defined in (ii) above, and the
third gene may be a gene as defined in (i), (iv) or (v) above (and
where both the second and third genes are introduced on plasmid or
vector, the second gene may or may not be introduced on the same
plasmid or vector as the third gene); [0129] the first gene may be
a gene as defined in (v) above, and the second gene may be a gene
as defined in (iii) above, and the third gene may be a gene as
defined in (i), (iv) or (v) above; [0130] the first gene may be a
gene as defined in (v) above, and the second gene may be a gene as
defined in (iv) above, and the third gene may be a gene as defined
in (i), (iii), (iv) or (v) above; or [0131] the first gene may be a
gene as defined in (v) above, and the second gene may be a gene as
defined in (v) above, and the third gene may be a gene as defined
in (i), (iv) or (v) above.
[0132] Further combinations of possible genetic modifications will
be apparent to the skilled person, in light of the above
disclosure, when further genes (for example a fourth gene encoding
a third helper protein; a fifth gene encoding a fourth helper
protein, etc.) are to be over-expressed in the host cell of the
invention.
[0133] The skilled person can readily choose the most appropriate
and convenient method to achieve over-expression of one or more
helper proteins in a host cell. It will be appreciated that, in the
case that multiple helper proteins are over-expressed in the host
cell, at least one helper protein may or may not be over-expressed
by the introduction of an appropriate recombinant polynucleotide
sequence as discussed above, whereas at least one other helper
protein may or may not be over-expressed by a genetic modification
to the host cell to cause over-expression of the helper protein
from the endogenous gene that encodes it.
Helper Proteins
[0134] As discussed above, we have identified a series of proteins
(hereinafter "helper" proteins) that are over-expressed in a S.
cerevisiae strain identified as possessing increased production of
a recombinant protein. These over-expressed helper proteins have
all, individually, been previously identified.
[0135] The helper proteins identified include proteins that can be
categorised as follows--
(i) chaperones, (ii) proteins involved in disulphide bond
formation, (iii) proteins involved in the protein degradation
pathway, and (iv) HAC1 (encoded by a spliced or unspliced
polynucleotide).
[0136] These groups are individually described further below.
Chaperones
[0137] The class of proteins known as chaperones have been defined
by Hartl (1996, Nature, 381, 571-580) as a protein that binds to
and stabilises an otherwise unstable conformer of another protein
and, by controlled binding and release, facilitates its correct
fate in vivo, be it folding, oligomeric assembly, transport to a
particular subcellular compartment, or disposal by degradation.
[0138] For the purposes of the present invention, chaperones of
interest can be broadly split into the following three functional
sub-groups-- [0139] ER luminal localised chaperones; [0140]
Chaperones involved in cytoplasmic folding and maintenance of
proteins in a translocation competent state prior to translocation;
and [0141] Mitochondrial chaperone and translocation proteins
[0142] Each of these groups are discussed in more detail below.
ER Luminal Localised Chaperones
[0143] ER luminal localised chaperones, involved in "protein
folding" include DnaJ-like proteins (such as JEM1), Hsp70 family
member proteins (such as LHS1), SCJ1, KAR2, SIL1 and FKB2. A
detailed description of these proteins and their genes is given
separately below.
[0144] In one embodiment, the host cell may or may not be
genetically modified to cause over-expression of one, or more, of
the above ER luminal localised chaperones. For example, SCJ1 may or
may not be over-expressed. Alternatively, FKB2 may or may not be
over-expressed.
[0145] In another embodiment, the host cell may or may not be
genetically modified to cause over-expression of two of the above
ER luminal localised chaperones. For example, one of the following
combinations may or may not be chosen-- [0146] A DnaJ-like proteins
(such as JEM1) in combination with one of an Hsp70 family member
protein (such as LHS1), SCJ1, KAR2, SIL1 or FKB2; [0147] An Hsp70
family member protein (such as LHS1) in combination with one of
SCJ1, KAR2, SIL1 or FKB2; [0148] SCJ1 in combination with one of
KAR2, KU or FKB2; [0149] KAR2 in combination with one of SIL1 or
FKB2; or [0150] SIL1 in combination with FKB2.
[0151] In another embodiment, the host cell may or may not be
genetically modified to cause over-expression of three of the above
ER luminal localised chaperones. For example, one of the following
combinations may or may not be chosen--
JEM1, LHS1 and SCJ1; JEM1, LHS1 and KAR2; JEM1, LHS1 and SIL1;
JEM1, LHS1 and FKB2; JEM1, SCJ1 and KAR2; JEM1, SCJ1 and SIL1;
JEM1, SCJ1 and FKB2; JEM1, KAR2 and SIL1; JEM1, KAR2 and FKB2;
JEM1, SIL1 and FKB2; LHS1, SCJ1 and KAR2; LHS1, SCJ1 and SIL1;
LHS1, SCJ1 and FKB2; LHS1, KAR2 and SIL1 LHS1, KAR2 and FKB2; LHS1,
SIL1 and FKB2; SCJ1, KAR2 and SIL1; SCJ1, KAR2 and FKB2; SCJ1, SIL1
and FKB2; or KAR2, SIL1 and FKB2.
[0152] In one embodiment, the host cell may or may not be
genetically modified to cause over-expression of four of the above
ER luminal localised chaperones. For example, one of the following
combinations may or may not be chosen--
JEM1, LHS1, SCJ1 and KAR2; JEM1, LHS1, SCJ1 and SIL1; JEM1, LHS1,
SCJ1 and FKB2; JEM1, LHS1, KAR2 and SIL1; JEM1, LHS1, KAR2 and
FKB2; JEM1, LHS1, SIL1 and FKB2; JEM1, SCJ1, KAR2 and SIL1; JEM1,
SCJ1, KAR2 and FKB2; JEM1, SCJ1, SIL1 and FKB2; JEM1, KAR2, SIL1
and FKB2; LHS1, SCJ1, KAR2 and SIL1; LHS1, SCJ1, KAR2 and FKB2;
LHS1, SCJ1, SIL1 and FKB2; LHS1, KAR2, SIL1 and FKB2; or SCJ1,
KAR2, SIL1 and FKB2.
[0153] In another embodiment, the host cell may or may not be
genetically modified to cause over-expression of five of the above
ER luminal localised chaperones. For example, one of the following
combinations may or may not be chosen--
JEM1, LHS1, SCJ1, KAR2 and SIL1; JEM1, LHS1, SCJ1, KAR2 and FKB2;
JEM1, LHS1, SCJ1, SIL1 and FKB2; JEM1, LHS1, KAR2, SIL1 and FKB2;
JEM1, SCJ1, KAR2, SIL1 and FKB2; or LHS1, SCJ1, KAR2, SIL1 and
FKB2.
[0154] In another embodiment, the host cell may or may not be
genetically modified to cause over-expression of all six of the
above ER luminal localised chaperones. In other words, the
following combination may or may not be chosen--
JEM1, LHS1, SCJ1, KAR2, SIL1 and FKB2.
[0155] In one preferred embodiment, the host cell may or may not be
genetically modified to cause over-expression of two, three or four
helper proteins selected from LHS1, SELL JEM1 and SCJ1, such as one
of the following combinations--
LHS1 and SIL1 LHS1 and JEM1; LHS1 and SCJ1; SIL1 and JEM1; SIL1 and
SCJ1; JEM1 and SCJ1; LHS1, SIL1 and JEM1; LHS1, SIL1 and SCJ1;
LHS1, JEM1 and SCJ1; SIL1 JEM1 and SCJ1; or LHS1, SIL1 JEM1 and
SCJ1.
Chaperones Involved in Cytoplasmic Folding and Maintenance of
Proteins in a Translocation Competent State Prior to
Translocation
[0156] Chaperones involved in cytoplasmic folding and maintenance
of proteins in a translocation competent state prior to
translocation include SSA1, SSA2, SSA3, SSA4, SSE1, SSE2, SSB1,
SSB2. A detailed description of these proteins and their genes is
given separately below.
[0157] In one embodiment, the host cell may or may not be
genetically modified to cause over-expression of one of the above
chaperones involved in cytoplasmic folding and maintenance of
proteins in a translocation competent state prior to translocation.
For example, SSE1 may or may not be chosen.
[0158] In another embodiment, the host cell may or may not be
genetically modified to cause over-expression of two of the above
chaperones involved in cytoplasmic folding and maintenance of
proteins in a translocation competent state prior to translocation.
For example, one of the following combinations may or may not be
chosen--
SSA1 in combination with one of SSA2, SSA3, SSA4, SSE1, SSE2, SSB1,
SSB2; SSA2 in combination with one of SSA3, SSA4, SSE1, SSE2, SSB1,
SSB2; SSA3 in combination with one of SSA4, SSE1, SSE2, SSB1, SSB2;
SSA4 in combination with one of SSE1, SSE2, SSB1, SSB2; SSE1 in
combination with one of SSE2, SSB1, SSB2; SSE2 in combination with
one of SSB1, SSB2; or SSB1 in combination with SSB2.
[0159] In another embodiment, the host cell may or may not be
genetically modified to cause over-expression of three of the above
chaperones involved in cytoplasmic folding and maintenance of
proteins in a translocation competent state prior to translocation.
For example, one of the following combinations may or may not be
chosen--
SSA1, SSA2 and SSA3; SSA1, SSA2 and SSA4; SSA1, SSA2 and SSE1;
SSA1, SSA2 and SSE2; SSA1, SSA2 and SSB1; SSA1, SSA2 and SSB2;
SSA1, SSA3 and SSA4; SSA1, SSA3 and SSE1; SSA1, SSA3 and SSE2;
SSA1, SSA3 and SSB1; SSA1, SSA3 and SSB2; SSA1, SSA4 and SSE1;
SSA1, SSA4 and SSE2; SSA1, SSA4 and SSB1; SSA1, SSA4 and SSB2;
SSA1, SSE1 and SSE2; SSA1, SSE1 and SSB1; SSA1, SSE1 and SSB2;
SSA1, SSE2 and SSB1; SSA1, SSE2 and SSB2; SSA1, SSB1 and SSB2;
SSA2, SSA3 and SSA4; SSA2, SSA3 and SSE1; SSA2, SSA3 and SSE2;
SSA2, SSA3 and SSB1; SSA2, SSA3 and SSB2; SSA2, SSA4 and SSE1;
SSA2, SSA4 and SSE2; SSA2, SSA4 and SSB1; SSA2, SSA4 and SSB2;
SSA2, SSE1 and SSE2; SSA2, SSE1 and SSB1; SSA2, SSE1 and SSB2;
SSA2, SSE2 and SSB1; SSA2, SSE2 and SSB2; SSA2, SSB1 and SSB2;
SSA3, SSA4 and SSE1; SSA3, SSA4 and SSE2; SSA3, SSA4 and SSB1;
SSA3, SSA4 and SSB2; SSA3, SSE1 and SSE2; SSA3, SSE1 and SSB1;
SSA3, SSE1 and SSB2; SSA3, SSE2 and SSB1; SSA3, SSE2 and SSB2;
SSA3, SSB1 and SSB2; SSA4, SSE1 and SSE2; SSA4, SSE1 and SSB1;
SSA4, SSE1 and SSB2; SSA4, SSE2 and SSB1; SSA4, SSE2 and SSB2;
SSA4, SSB1 and SSB2; SSE1, SSE2 and SSB1; SSE1, SSE2 and SSB2;
SSE1, SSB1 and SSB2; or SSE2, SSB1 and SSB2.
[0160] In another embodiment, the host cell mayor may not be
genetically modified to cause over-expression of four of the above
chaperones involved in cytoplasmic folding and maintenance of
proteins in a translocation competent state prior to translocation.
For example, one of the following combinations may or may not be
chosen--
SSA1, SSA2, SSA3 and SSA4; SSA1, SSA2, SSA3 and SSE1; SSA1, SSA2,
SSA3 and SSE2; SSA1, SSA2, SSA3 and SSB1; SSA1, SSA2, SSA3 and
SSB2; SSA1, SSA2, SSA4 and SSE1; SSA1, SSA2, SSA4 and SSE2; SSA1,
SSA2, SSA4 and SSB1; SSA1, SSA2, SSA4 and SSB2; SSA1, SSA2, SSE1
and SSE2; SSA1, SSA2, SSE1 and SSB1; SSA1, SSA2, SSE1 and SSB2;
SSA1, SSA2, SSE2 and SSB1; SSA1, SSA2, SSE2 and SSB2; SSA1, SSA2,
SSB1 and SSB2; SSA1, SSA3, SSA4 and SSE1; SSA1, SSA3, SSA4 and
SSE2; SSA1, SSA3, SSA4 and SSB1; SSA1, SSA3, SSA4 and SSB2; SSA1,
SSA3, SSE1 and SSE2; SSA1, SSA3, SSE1 and SSB1; SSA1, SSA3, SSE1
and SSB2; SSA1, SSA3, SSE2 and SSB1; SSA1, SSA3, SSE2 and SSB2;
SSA1, SSA3, SSB1 and SSB2; SSA1, SSA4, SSE1 and SSE2; SSA1, SSA4,
SSE1 and SSB1; SSA1, SSA4, SSE1 and SSB2; SSA1, SSA4, SSE2 and
SSB1; SSA1, SSA4, SSE2 and SSB2; SSA1, SSA4, SSB1 and SSB2; SSA1,
SSE1, SSE2 and SSB1; SSA1, SSE1, SSE2 and SSB2; SSA1, SSE1, SSB1
and SSB2; SSA1, SSE2, SSB1 and SSB2; SSA2, SSA3, SSA4 and SSE1;
SSA2, SSA3, SSA4 and SSE2; SSA2, SSA3, SSA4 and SSB1; SSA2, SSA3,
SSA4 and SSB2; SSA2, SSA3, SSE1 and SSE2; SSA2, SSA3, SSE1 and
SSB1; SSA2, SSA3, SSE1 and SSB2; SSA2, SSA3, SSE2 and SSB1; SSA2,
SSA3, SSE2 and SSB2; SSA2, SSA3, SSB1 and SSB2; SSA2, SSA4, SSE1
and SSE2; SSA2, SSA4, SSE1 and SSB1; SSA2, SSA4, SSE1 and SSB2;
SSA2, SSA4, SSE2 and SSB1; SSA2, SSA4, SSE2 and SSB2; SSA2, SSA4,
SSB1 and SSB2; SSA2, SSE1, SSE2 and SSB1; SSA2, SSE1, SSE2 and
SSB2; SSA2, SSE1, SSB1 and SSB2; SSA2, SSE2, SSB1 and SSB2; SSA3,
SSA4, SSE1 and SSE2; SSA3, SSA4, SSE1 and SSB1; SSA3, SSA4, SSE1
and SSB2; SSA3, SSA4, SSE2 and SSB1; SSA3, SSA4, SSE2 and SSB2;
SSA3, SSA4, SSB1 and SSB2; SSA3, SSE1, SSE2 and SSB1; SSA3, SSE1,
SSE2 and SSB2; SSA3, SSE1, SSB1 and SSB2; SSA3, SSE2, SSB1 and
SSB2; SSA4, SSE1, SSE2 and SSB1; SSA4, SSE1, SSE2 and SSB2; SSA4,
SSE1, SSB1 and SSB2; SSA4, SSE2, SSB1 and SSB2; or SSE1, SSE2, SSB1
and SSB2.
[0161] In another embodiment, the host cell may or may not be
genetically modified to cause over-expression of five of the above
chaperones involved in cytoplasmic folding and maintenance of
proteins in a translocation competent state prior to translocation.
For example, one of the following combinations may or may not be
chosen--
SSA1, SSA2, SSA3, SSA4 and SSE1; SSA1, SSA2, SSA3, SSA4 and SSE2;
SSA1, SSA2, SSA3, SSA4 and SSB1; SSA1, SSA2, SSA3, SSA4 and SSB2;
SSA1, SSA2, SSA3, SSE1 and SSE2; SSA1, SSA2, SSA3, SSE1 and SSB1;
SSA1, SSA2, SSA3, SSE1 and SSB2; SSA1, SSA2, SSA3, SSE2 and SSB1;
SSA1, SSA2, SSA3, SSE2 and SSB2; SSA1, SSA2, SSA3, SSB1 and SSB2;
SSA1, SSA2, SSA4, SSE1 and SSE2; SSA1, SSA2, SSA4, SSE1 and SSB1;
SSA1, SSA2, SSA4, SSE1 and SSB2; SSA1, SSA2, SSA4, SSE2 and SSB1;
SSA1, SSA2, SSA4, SSE2 and SSB2; SSA1, SSA2, SSA4, SSB1 and SSB2;
SSA1, SSA2, SSE1, SSE2 and SSB1; SSA1, SSA2, SSE1, SSE2 and SSB2;
SSA1, SSA2, SSE1, SSB1 and SSB2; SSA1, SSA2, SSE2, SSB1 and SSB2;
SSA1, SSA3, SSA4, SSE1 and SSE2; SSA1, SSA3, SSA4, SSE1 and SSB1;
SSA1, SSA3, SSA4, SSE1 and SSB2; SSA1, SSA3, SSA4, SSE2 and SSB1;
SSA1, SSA3, SSA4, SSE2 and SSB2; SSA1, SSA3, SSA4, SSB1 and SSB2;
SSA1, SSA3, SSE1, SSE2 and SSB1; SSA1, SSA3, SSE1, SSE2 and SSB2;
SSA1, SSA3, SSE1, SSB1 and SSB2; SSA1, SSA3, SSE2, SSB1 and SSB2;
SSA1, SSA4, SSE1, SSE2 and SSB1; SSA1, SSA4, SSE1, SSE2 and SSB2;
SSA1, SSA4, SSE1, SSB1 and SSB2; SSA1, SSE1, SSE2, SSB1 and SSB2;
SSA2, SSA3, SSA4, SSE1 and SSE2; SSA2, SSA3, SSA4, SSE1 and SSB1;
SSA2, SSA3, SSA4, SSE1 and SSB2; SSA2, SSA3, SSA4, SSE2 and SSB1;
SSA2, SSA3, SSA4, SSE2 and SSB2; SSA2, SSA3, SSA4, SSB1 and SSB2;
SSA2, SSA3, SSE1, SSE2 and SSB1; SSA2, SSA3, SSE1, SSE2 and SSB2;
SSA2, SSA3, SSE1, SSB1 and SSB2; SSA2, SSA3, SSE2, SSB1 and SSB2;
SSA2, SSA4, SSE1, SSE2 and SSB1; SSA2, SSA4, SSE1, SSE2 and SSB2;
SSA2, SSA4, SSE1, SSB1 and SSB2; SSA2, SSA4, SSE2, SSB1 and SSB2;
SSA2, SSE1, SSE2, SSB1 and SSB2; SSA3, SSA4, SSE1, SSE2 and SSB1;
SSA3, SSA4, SSE1, SSE2 and SSB2; SSA3, SSA4, SSE1, SSB1 and SSB2;
SSA3, SSA4, SSE2, SSB1 and SSB2; SSA3, SSE1, SSE2, SSB1 and SSB2;
or SSA4, SSE1, SSE2, SSB1 and SSB2.
[0162] In another embodiment, the host cell may or may not be
genetically modified to cause over-expression of six of the above
chaperones involved in cytoplasmic folding and maintenance of
proteins in a translocation competent state prior to translocation.
For example, one of the following combinations may or may not be
chosen--
SSA1, SSA2, SSA3, SSA4, SSE1 and SSE2; SSA1, SSA2, SSA3, SSA4, SSE1
and SSB1; SSA1, SSA2, SSA3, SSA4, SSE1 and SSB2; SSA1, SSA2, SSA3,
SSA4, SSE2 and SSB1; SSA1, SSA2, SSA3, SSA4, SSE2 and SSB2; SSA1,
SSA2, SSA3, SSA4, SSB1 and SSB2; SSA1, SSA2, SSA3, SSE1, SSE2 and
SSB1; SSA1, SSA2, SSA3, SSE1, SSE2 and SSB2; SSA1, SSA2, SSA3,
SSE1, SSB1 and SSB2; SSA1, SSA2, SSA3, SSE2, SSB1 and SSB2; SSA1,
SSA2, SSA4, SSE1, SSE2 and SSB1; SSA1, SSA2, SSA4, SSE1, SSE2 and
SSB2; SSA1, SSA2, SSA4, SSE1, SSB1 and SSB2; SSA1, SSA2, SSA4,
SSE2, SSB1 and SSB2; SSA1, SSA2, SSE1, SSE2, SSB1 and SSB2; SSA1,
SSA3, SSA4, SSE1, SSE2 and SSB1; SSA1, SSA3, SSA4, SSE1, SSE2 and
SSB2; SSA1, SSA3, SSA4, SSE1, SSB1 and SSB2; SSA1, SSA3, SSA4,
SSE2, SSB1 and SSB2; SSA1, SSA3, SSE1, SSE2, SSB1 and SSB2; SSA1,
SSA4, SSE1, SSE2, SSB1 and SSB2; SSA2, SSA3, SSA4, SSE1, SSE2 and
SSB1; SSA2, SSA3, SSA4, SSE1, SSE2 and SSB2; SSA2, SSA3, SSA4,
SSE1, SSB1 and SSB2; SSA2, SSA3, SSA4, SSE2, SSB1 and SSB2; SSA2,
SSA3, SSE1, SSE2, SSB1 and SSB2; SSA2, SSA4, SSE1, SSE2, SSB1 and
SSB2; or SSA3, SSA4, SSE1, SSE2, SSB1 and SSB2.
[0163] In another embodiment, the host cell may or may not be
genetically modified to cause over-expression of seven of the above
chaperones involved in cytoplasmic folding and maintenance of
proteins in a translocation competent state prior to translocation.
For example, one of the following combinations may or may not be
chosen--
SSA1, SSA2, SSA3, SSA4, SSE1, SSE2 and SSB1; SSA1, SSA2, SSA3,
SSA4, SSE1, SSE2 and SSB; SSA1, SSA2, SSA3, SSA4, SSE1, SSB1 and
SSB2; SSA1, SSA2; SSA3, SSA4, SSE2, SSB1 and SSB2; SSA1, SSA2,
SSA3, SSE1, SSE2, SSB1 and SSB2; SSA1, SSA2, SSA4, SSE1, SSE2, SSB1
and SSB2; SSA1, SSA3, SSA4, SSE1, SSE2, SSB1 and SSB2; or SSA2,
SSA3, SSA4, SSE1, SSE2, SSB1 and SSB2.
[0164] In another embodiment, the host cell may or may not be
genetically modified to cause over-expression of all eight of the
above chaperones involved in cytoplasmic folding and maintenance of
proteins in a translocation competent state prior to translocation.
In other words, the following combination may or may not be
chosen--
SSA1, SSA2, SSA3, SSA4, SSE1, SSE2, SSB1 and SSB2.
Mitochondrial Chaperone and Translocation Proteins
[0165] Mitochondrial chaperone and translocation proteins include
ECM10, MDJ1, MDJ2. A detailed description of these proteins and
their genes is given separately below.
[0166] In one embodiment, the host cell may or may not be
genetically modified to cause over-expression of one of the above
mitochondrial chaperone and translocation proteins.
[0167] In another embodiment, the host cell may or may not be
genetically modified to cause over-expression of two of the above
mitochondrial chaperone and translocation proteins. For example,
one of the following combinations may or may not be chosen--
ECM10 and MDJ1; ECM10 and MDJ2; or MDJ1 and MDJ2.
[0168] In another embodiment, the host cell may or may not be
genetically modified to cause over-expression of all three of the
above mitochondrial chaperone and translocation proteins. In that
case the following combination may or may not be chosen--
ECM10, MDJ1 and MDJ2.
Other Combinations of Chaperones
[0169] The skilled person will appreciate that it is possible to
combine genes that encode one or more proteins from the
above-defined groups of chaperones.
[0170] Thus, the host cell may or may not be genetically modified
to cause simultaneous over-expression of at least one, two, three,
four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen,
fourteen, fifteen, sixteen or seventeen of the chaperones selected
from the group consisting of JEM1, LHS1, SCJ1, KAR2, SIL1 FKB2,
SSA1, SSA2, SSA3, SSA4, SSE1, SSE2, SSB1, SSB2, ECM10, MDJ1 and
MDJ2.
[0171] Where the host cell is genetically modified to cause
simultaneous over-expression of one or two of the above defined
chaperones, it may or may not be preferred that the host cell is
genetically modified to cause simultaneous over-expression of at
least three helper proteins and the one or two other helper
proteins may or may not be helper proteins involved in disulphide
bond formation or protein degradation, as discussed below.
[0172] Over-expression of one (or more) of the ER luminal localised
chaperones may or may not be combined with the over-expression of
at least one of the chaperones involved in cytoplasmic folding and
maintenance of proteins in a translocation competent state prior to
translocation and/or the over-expression of at least one of the
mitochondrial chaperone and translocation proteins.
[0173] For example, any one of the following combinations may or
may not be chosen-- [0174] SCJ1 in combination with any one, two,
three, four, five, six, seven, eight, nine, ten or eleven of SSA1,
SSA2, SSA3, SSA4, SSE1, SSE2, SSB1, SSB2, ECM10, MDJ1 or MDJ2; or
[0175] FKB2 in combination with any one, two, three, four, five,
six, seven, eight, nine, ten or eleven of SSA1, SSA2, SSA3, SSA4,
SSE1, SSE2, SSB1, SSB2, ECM10, MDJ1 or MDJ2. [0176] JEM1 in
combination with any of the above-listed combinations of one, two,
three, four, five, six, seven or eight of SSA1, SSA2, SSA3, SSA4,
SSE1, SSE2, SSB1, SSB2 and/or in combination with ECM10, MDJ1 and
MDJ2; [0177] LHS1 in combination with any of the above-listed
combinations of one, two, three, four, five, six, seven or eight of
SSA1, SSA2, SSA3, SSA4, SSE1, SSE2, SSB1, SSB2 and/or in
combination with ECM10, MDJ1 and MDJ2; [0178] SCJ1 in combination
with any of the above-listed combinations of one, two, three, four,
five, six, seven or eight of SSA1, SSA2, SSA3, SSA4, SSE1, SSE2,
SSB1, SSB2 and/or in combination with ECM10, MDJ1 and MDJ2; [0179]
KAR2 in combination with any of the above-listed combinations of
one, two, three, four, five, six, seven or eight of SSA1, SSA2,
SSA3, SSA4, SSE1, SSE2, SSB1, SSB2 and/or in combination with
ECM10, MDJ1 and MDJ2; [0180] SIL1 in combination with any of the
above-listed combinations of one, two, three, four, five, six,
seven or eight of SSA1, SSA2, SSA3, SSA4, SSE1, SSE2, SSB1, SSB2
and/or in combination with ECM10, MDJ1 and MDJ2; or [0181] FKB2 in
combination with any of the above-listed combinations of one, two,
three, four, five, six, seven or eight of SSA1, SSA2, SSA3, SSA4,
SSE1, SSE2, SSB1, SSB2 and/or in combination with ECM10, MDJ1 and
MDJ2.
[0182] Alternatively, for example, one (or more) of the chaperones
involved in cytoplasmic folding and maintenance of proteins in a
translocation competent state prior to translocation may or may not
be simultaneously over-expressed with at least one of the ER
luminal localised chaperones and/or at least one of the
mitochondrial chaperone and translocation proteins. For example,
the following combinations may or may not be chosen-- [0183] SSE1
in combination with any one, two, three, four, five, six, seven,
eight or nine of JEM1, LHS1, SCJ1, KAR2, SIL1, FKB2, ECM10, MDJ1 or
MDJ2. [0184] SSA1 in combination with any of the above-listed
combinations of one, two, three, four, five or six of JEM1, LHS1,
SCJ1, KAR2, SIL1 and FKB2 and/or in combination with ECM10, MDJ1
and MDJ2; [0185] SSA2 in combination with any of the above-listed
combinations of one, two, three, four, five or six of JEM1, LHS1,
SCJ1, KAR2, SIL1 and FKB2 and/or in combination with ECM10, MDJ1
and MDJ2; [0186] SSA3 in combination with any of the above-listed
combinations of one, two, three, four, five or six of JEM1, LHS1,
SCJ1, KAR2, SIL1 and FKB2 and/or in combination with ECM10, MDJ1
and MDJ2; [0187] SSA4 in combination with any of the above-listed
combinations of one, two, three, four, five or six of JEM1, LHS1,
SCJ1, KAR2, SIL1 and FKB2 and/or in combination with ECM10, MDJ1
and MDJ2; [0188] SSE1 in combination with any of the above-listed
combinations of one, two, three, four, five or six of JEM1, LHS1,
SCJ1, KAR2, SIL1 and FKB2 and/or in combination with ECM10, MDJ1
and MDJ2; [0189] SSE2 in combination with any of the above-listed
combinations of one, two, three, four, five or six of JEM1, LHS1,
SCJ1, KAR2, SIL1 and FKB2 and/or in combination with ECM10, MDJ1
and MDJ2; [0190] SSB1 in combination with any of the above-listed
combinations of one, two, three, four, five or six of JEM1, LHS1,
SCJ1, KAR2, SIL1 and FKB2 and/or in combination with ECM10, MDJ1
and MDJ2; or [0191] SSB2 in combination with any of the
above-listed combinations of one, two, three, four, five or six of
JEM1, LHS1, SCJ1, KAR2, SIL1 and FKB2 and/or in combination with
ECM10, MDJ1 and MDJ2.
[0192] Alternatively, one of the mitochondrial chaperone and
translocation proteins may or may not be simultaneously
over-expressed with at least one of the chaperones involved in
cytoplasmic folding and maintenance of proteins in a translocation
competent state prior to translocation and/or at least one of the
ER luminal localised chaperones.
[0193] For example, one of the following combinations may or may
not be chosen-- [0194] ECM10 in combination with any of the
above-listed combinations of one, two, three, four, five or six of
JEM1, LHS1, SCJ1, KAR2, SIL1 and FKB2; [0195] ECM10 in combination
with any of the above-listed combinations of one, two, three, four,
five, six, seven or eight of SSA1, SSA2, SSA3, SSA4, SSE1, SSE2,
SSB1, SSB2; [0196] ECM10 in combination with any of the
above-listed combinations of one, two, three of JEM1, LHS1, SCJ1,
KAR2, SIL1 and FKB2 and any of the above-listed combinations of
one, two, three, four, five, six, seven or eight of SSA1, SSA2,
SSA3, SSA4, SSE1, SSE2, SSB1, SSB2; [0197] ECM10 in combination
with any of the above-listed combinations of four of JEM1, LHS1,
SCJ1, KAR2, SIL1 and FKB2 and any of the above-listed combinations
of one, two, three, four, five, six, seven or eight of SSA1, SSA2,
SSA3, SSA4, SSE1, SSE2, SSB1, SSB2; [0198] ECM10 in combination
with any of the above-listed combinations of five of JEM1, LHS1,
SCJ1, KAR2, SILT and FKB2 and any of the above-listed combinations
of one, two, three, four, five, six, seven or eight of SSA1, SSA2,
SSA3, SSA4, SSE1, SSE2, SSB1, SSB2; [0199] ECM10 in combination
with all six of JEM1, LHS1, SCJ1, KAR2, SIL1 and FKB2 and any of
the above-listed combinations of one, two, three, four, five, six,
seven or eight of SSA1, SSA2, SSA3, SSA4, SSE1, SSE2, SSB1, SSB2;
[0200] MDJ1 in combination with any of the above-listed
combinations of one, two, three, four, five or six of JEM1, LHS1,
--SCJ1, KAR2, SIL1 and FKB2; [0201] MDJ1 in combination with any of
the above-listed combinations of one, two, three, four, five, six,
seven or eight of SSA1, SSA2, SSA3, SSA4, SSE1, SSE2, SSB1, SSB2;
[0202] MDJ1 in combination with any of the above-listed
combinations of one, two, three of JEM1, LHS1, SCJ1, KAR2, SIL1 and
FKB2 and any of the above-listed combinations of one, two, three,
four, five, six, seven or eight of SSA1, SSA2, SSA3, SSA4, SSE1,
SSE2, SSB1, SSB2; [0203] MDJ1 in combination with any of the
above-listed combinations of four of JEM1, LHS1, SCJ1, KAR2, SIL1
and FKB2 and any of the above-listed combinations of one, two,
three, four, five, six, seven or eight of SSA1, SSA2, SSA3, SSA4,
SSE1, SSE2, SSB1, SSB2; [0204] MDJ1 in combination with any of the
above-listed combinations of five of JEM1, LHS1, SCJ1, KAR2, SIL1
and FKB2 and any of the above-listed combinations of one, two,
three, four, five, six, seven or eight of SSA1, SSA2, SSA3, SSA4,
SSE1, SSE2, SSB1, SSB2; [0205] MDJ1 in combination with all six of
JEM1, LHS1, SCJ1, KAR2, SIL1 and FKB2 and any of the above-listed
combinations of one, two, three, four, five, six, seven or eight of
SSA1, SSA2, SSA3, SSA4, SSE1, SSE2, SSB1, SSB2; [0206] MDJ2 in
combination with any of the above-listed combinations of one, two,
three, four, five or six of JEM1, LHS1, SCJ1, KAR2, SIL1 and FKB2;
[0207] MDJ2 in combination with any of the above-listed
combinations of one, two, three, four, five, six, seven or eight of
SSA1, SSA2, SSA3, SSA4, SSE1, SSE2, SSB1, SSB2; [0208] MDJ2 in
combination with any of the above-listed combinations of one, two,
three of JEM1, LHS1, SCJ1, KAR2, SIL1 and FKB2 and any of the
above-listed combinations of one, two, three, four, five, six,
seven or eight of SSA1, SSA2, SSA3, SSA4, SSE1, SSE2, SSB1, SSB2;
[0209] MDJ2 in combination with any of the above-listed
combinations of four of JEM1, LHS1, SCJ1, KAR2, SIL1 and FKB2 and
any of the above-listed combinations of one, two, three, four,
five, six, seven or eight of SSA1, SSA2, SSA3, SSA4, SSE1, SSE2,
SSB1, SSB2; [0210] MDJ2 in combination with any of the above-listed
combinations of five of JEM1, LHS1, SCJ1, KAR2, SIL1 and FKB2 and
any of the above-listed combinations of one, two, three, four,
five, six, seven or eight of SSA1, SSA2, SSA3, SSA4, SSE1, SSE2,
SSB1, SSB2; or [0211] MDJ2 in combination with all six of JEM1,
LHS1, SCJ1, KAR2, SIL1 and FKB2 and any of the above-listed
combinations of one, two, three, four, five, six, seven or eight of
SSA1, SSA2, SSA3, SSA4, SSE1, SSE2, SSB1, SSB2.
[0212] In another embodiment, representative members of each of the
above three groups of chaperone proteins (such as one member of
each group) may or may not be simultaneously over-expressed in the
host cell. For example, one of the following combinations may or
may not be chosen--
JEM1, SSA1 and ERM10; JEM1, SSA1 and MDJ1; JEM1, SSA1 and MDJ2;
JEM1, SSA2 and ERM10; JEM1, SSA2 and MDJ1; JEM1, SSA2 and MDJ2;
JEM1, SSA3 and ERM10; JEM1, SSA3 and MDJ1; JEM1, SSA3 and MDJ2;
JEM1, SSA4 and ERM10; JEM1, SSA4 and MDJ1; JEM1, SSA4 and MDJ2;
JEM1, SSE1 and ERM10; JEM1, SSE1 and MDJ1; JEM1, SSE1 and MDJ2;
JEM1, SSE2 and ERM10; JEM1, SSE2 and MDJ1; JEM1, SSE2 and MDJ2;
JEM1, SSB1 and ERM10; JEM1, SSB1 and MDJ1; JEM1, SSB1 and MDJ2;
JEM1, SSB2 and ERM10; JEM1, SSB2 and MDJ1; JEM1, SSB2 and MDJ2;
LHS1, SSA1 and ERM10; LHS1, SSA1 and MDJ1; LHS1, SSA1 and MDJ2;
LHS1, SSA2 and ERM10; LHS1, SSA2 and MDJ1; LHS1, SSA2 and MDJ2;
LHS1, SSA3 and ERM10; LHS1, SSA3 and MDJ1 LHS1, SSA3 and MDJ2;
LHS1, SSA4 and ERM10; LHS1, SSA4 and MDJ1; LHS1, SSA4 and MDJ2;
LHS1, SSE1 and ERM10; LHS1, SSE1 and MDJ1; LHS1, SSE1 and MDJ2;
LHS1, SSE2 and ERM10; LHS1, SSE2 and MDJ1; LHS1, SSE2 and MDJ2;
LHS1, SSB1 and ERM10; LHS1, SSB1 and MDJ1; LHS1, SSB1 and MDJ2;
LHS1, SSB2 and ERM10; LHS1, SSB2 and MDJ1; LHS1, SSB2 and MDJ2;
SCJ1, SSA1 and ERM10; SCJ1, SSA1 and MDJ1; SCJ1, SSA1 and MDJ2;
SCJ1, SSA2 and ERM10; SCJ1, SSA2 and MDJ1; SCJ1, SSA2 and MDJ2;
SCJ1, SSA3 and ERM10; SCJ1, SSA3 and MDJ1; SCJ1, SSA3 and MDJ2;
SCJ1, SSA4 and ERM10; SCJ1, SSA4 and MDJ1; SCJ1, SSA4 and MDJ2;
SCJ1, SSE1 and ERM10; SCJ1, SSE1 and MDJ1; SCJ1, SSE1 and MDJ2;
SCJ1, SSE2 and ERM10; SCJ1, SSE2 and MDJ1; SCJ1, SSE2 and MDJ2;
SCJ1, SSB1 and ERM10; SCJ1, SSB1 and MDJ1; SCJ1, SSB1 and MDJ2;
SCJ1, SSB2 and ERM10; SCJ1, SSB2 and MDJ1; SCJ1, SSB2 and MDJ2;
KAR2, SSA1 and ERM10; KAR2, SSA1 and MDJ1; KAR2, SSA1 and MDJ2;
KAR2, SSA2 and ERM10; KAR2, SSA2 and MDJ1; KAR2, SSA2 and MDJ2;
KAR2, SSA3 and ERM10; KAR2, SSA3 and MDJ1; KAR2, SSA3 and MDJ2;
KAR2, SSA4 and ERM10; KAR2, SSA4 and MDJ1; KAR2, SSA4 and MDJ2;
KAR2, SSE1 and ERM10; KAR2, SSE1 and MDJ1; KAR2, SSE1 and MDJ2;
KAR2, SSE2 and ERM10; KAR2, SSE2 and MDJ1; KAR2, SSE2 and MDJ2;
KAR2, SSB1 and ERM10; KAR2, SSB1 and MDJ1; KAR2, SSB1 and MDJ2;
KAR2, SSB2 and ERM10; KAR2, SSB2 and MDJ1; KAR2, SSB2 and MDJ2;
SIL1 SSA1 and ERM10; SIL1 SSA1 and MDJ1; SIL1 SSA1 and MDJ2; SSA2
and ERM10; SIL1 SSA2 and MDJ1; SIL1 SSA2 and MDJ2; SIL1, SSA3 and
ERM10; SIL1 SSA3 and MDJ1; SIL1 SSA3 and MDJ2; SIL1 SSA4 and ERM10;
SIL1 SSA4 and MDJ1; SIL1 SSA4 and MDJ2; SIL1, SSE1 and ERM10; SIL1,
SSE1 and MDJ1; SIL1, SSE1 and MDJ2; SIL1, SSE2 and ERM10; SIL1,
SSE2 and MDJ1; SIL1, SSE2 and MDJ2; SIL1, SSB1 and ERM10; SIL1,
SSB1 and MDJ1; SIL1, SSB1 and MDJ2; SIL1, SSB2 and ERM10; SIL1,
SSB2 and MDJ1; SIL1, SSB2 and MDJ2; FKB2, SSA1 and ERM10; FKB2,
SSA1 and MDJ1; FKB2, SSA1 and MDJ2; FKB2, SSA2 and ERM10; FKB2,
SSA2 and MDJ1; FKB2, SSA2 and MDJ2; FKB2, SSA3 and ERM10; FKB2,
SSA3 and MDJ1; FKB2, SSA3 and MDJ2; FKB2, SSA4 and ERM10; FKB2,
SSA4 and MDJ1; FKB2, SSA4 and MDJ2; FKB2, SSE1 and ERM10; FKB2,
SSE1 and MDJ1; FKB2, SSE1 and MDJ2; FKB2, SSE2 and ERM10; FKB2,
SSE2 and MDJ1; FKB2, SSE2 and MDJ2; FKB2, SSB1 and ERM10; FKB2,
SSB1 and MDJ1; FKB2, SSB1 and MDJ2; FKB2, SSB2 and ERM10; FKB2,
SSB2 and MDJ1; or FKB2, SSB2 and MDJ2.
[0213] The skilled person will also appreciate that any of the
above defined combinations may or may not also be combined with any
of the following genes or combinations of genes encoding other
helper proteins, in particular helper proteins involved in
disulphide bond formation or helper proteins involved in protein
degradation, as discussed below.
Proteins Involved in Disulphide Bond Formation
[0214] Proteins involved in the formation of disulphide bonds in
other proteins include ERO1, ERV2, EUG1, MPD1, MPD2, EPS1 and PDI1.
A detailed description of these proteins and their genes is given
separately below.
[0215] In one embodiment, one of the above disulphide bond
formation proteins may or may not be over-expressed in the host
cell. For example, ERV2 may or may not be chosen.
[0216] In another embodiment, two of the above disulphide bond
formation proteins may or may not be simultaneously over-expressed
in the host cell. For example, one of the following combinations
may or may not be chosen-- [0217] ERO1 in combination with one of
ERV2, EUG1, MPD1, MPD2, EPS1 or PDI1; [0218] ERV2 in combination
with one of EUG1, MPD1, MPD2, EPS1 or PDI1; [0219] EUG1 in
combination with one of MPD1, MPD2, EPS1 or PDI1; [0220] MPD1 in
combination with one of MPD2, EPS1 or PDI1; [0221] MPD2 in
combination with one of EPS1 or PDI1; or [0222] EPS1 in combination
with PDI1.
[0223] In another embodiment, three of the above helper proteins
may or may not be simultaneously over-expressed in the host cell.
For example, one of the following combinations may or may not be
chosen--
ERO1, ERV2 and EUG1; ERO1, ERV2 and MPD1; ERO1, ERV2 and MPD2;
ERO1, ERV2 and EPS1; ERO1, ERV2 and PDI1; ERO1, EUG1 and MPD1;
ERO1, EUG1 and MPD2; ERO1, EUG1 and EPS1; ERO1, EUG1 and PDI1;
ERO1, MPD1 and MPD2; ERO1, MPD1 and EPS1; ERO1, MPD1 and PDI1;
ERO1, MPD2 and EPS1; ERO1, MPD2 and PDI1; ERO1, EPS1 and PDI1;
ERV2, EUG1 and MPD1; ERV2, EUG1 and MPD2; ERV2, EUG1 and EPS1;
ERV2, EUG1 and PDI1; ERV2, MPD1 and MPD2; ERV2, MPD1 and EPS1;
ERV2, MPD1 and PDI1; ERV2, MPD2 and EPS1; ERV2, MPD2 and PDI1;
ERV2, EPS1 and PDI1; EUG1, MPD1 and MPD2; EUG1, MPD1 and EPS1;
EUG1, MPD1 and PDI1; EUG1, MPD2 and EPS1; EUG1, MPD2 and PDI1;
EUG1, EPS1 and PDI1; MPD1, MPD2 and EPS1; MPD1, MPD2 and PDI1;
MPD1, EPS1 and PDI1; or MPD2; EPS1 and PDI1.
[0224] In another embodiment, four of the above helper proteins may
or may not be simultaneously over-expressed in the host cell. For
example, one of the following combinations may or may not be
chosen--
ERO1, ERV2, EUG1 and MPD1; ERO1, ERV2, EUG1 and MPD2; ERO1, ERV2,
EUG1 and EPS1; ERO1, ERV2, EUG1 and PDI1; ERO1, ERV2, MPD1 and
MPD2; ERO1, ERV2, MPD1 and EPS1; ERO1, ERV2, MPD1 and PDI1; ERO1,
ERV2, MPD2 and EPS1; ERO1, ERV2, MPD2 and PDI1; ERO1, ERV2, EPS1
and PDI1; ERO1, EUG1, MPD1 and MPD2; ERO1, EUG1, MPD1 and EPS1;
ERO1, EUG1, MPD1 and PDI1; ERO1, EUG1, MPD2 and EPS1; ERO1, EUG1,
MPD2 and PDI1; ERO1, EUG1, EPS1 and PDI1; ERO1, MPD1, MPD2 and
EPS1; ERO1, MPD1, MPD2 and PDI1; ERO1, MPD1, EPS1 and PDI1; ERO1,
MPD2, EPS1 and PDI1; ERV2, EUG1, MPD1 and MPD2; ERV2, EUG1, MPD1
and EPS1; ERV2, EUG1, MPD1 and PDI1; ERV2, EUG1, MPD2 and EPS1;
ERV2, EUG1, MPD2 and PDI1; ERV2, EUG1, EPS1 and PDI1; ERV2, MPD1,
MPD2 and EPS1; ERV2, MPD1, MPD2 and PDI1; ERV2, MPD1, EPS1 and
PDI1; ERV2, MPD2, EPS1 and PDI1; EUG1, MPD1, MPD2 and EPS1; EUG1,
MPD1, MPD2 and PDI1; EUG1, MPD1, EPS1 and PDI1; EUG1, MPD2, EPS1
and PDI1; or MPD1, MPD2, EPS1 and PDI1.
[0225] In another embodiment, five of the above helper proteins may
or may not be simultaneously over-expressed in the host cell. For
example, one of the following combinations may or may not be
chosen--
ERO1, ERV2, EUG1, MPD1 and MPD2; ERO1, ERV2, EUG1, MPD1 and EPS1;
ERO1, ERV2, EUG1, MPD1 and PDI1; ERO1, ERV2, EUG1, MPD2 and EPS1;
ERO1, ERV2, EUG1, MPD2 and PDI1; ERO1, ERV2, EUG1, EPS1 and PDI1;
ERO1, ERV2, MPD1, MPD2 and EPS1; ERO1, ERV2, MPD1, MPD2 and PDI1;
ERO1, ERV2, MPD1, EPS1 and PDI1; ERO1, ERV2, MPD2, EPS1 and PDI1;
ERO1, EUG1, MPD1, MPD2 and EPS1; ERO1, EUG1, MPD1, MPD2 and PDI1;
ERO1, EUG1, MPD1, EPS1 and PDI1; ERO1, EUG1, MPD2, EPS1 and PDI1;
ERO1, MPD1, MPD2, EPS1 and PDI1; ERV2, EUG1, MPD1, MPD2 and EPS1;
ERV2, EUG1, MPD1, MPD2 and PDI1; ERV2, EUG1, MPD1, EPS1 and PDI1;
ERV2, EUG1, MPD2, EPS1 and PDI1; ERV2, MPD1, MPD2, EPS1 and PDI1;
or EUG1, MPD1, MPD2, EPS1 and PDI1
[0226] In another embodiment, six of the above helper proteins may
or may not be simultaneously over-expressed in the host cell. For
example, one of the following combinations may or may not be
chosen--
ERO1, ERV2, EUG1, MPD1, MPD2 and EPS1; ERO1, ERV2, EUG1, MPD1, MPD2
and PDI1; ERO1, ERV2, EUG1, MPD1, EPS1 and PDI1; ERO1, ERV2, EUG1,
MPD2, EPS1 and PDI1; ERO1, ERV2, MPD1, MPD2, EPS1 and PDI1; ERO1,
EUG1, MPD1, MPD2, EPS1 and PDI1; or ERV2, EUG1, MPD1, MPD2, EPS1
and PDI1.
[0227] It is anticipated that ERO1 and ERV2 may function
independently of each other or they may co-operate. Therefore, in
one embodiment disclosure of ERO1 may or may not also include the
combinations of ERO1 and ERV2, or ERV2 in its place. Similarly, in
another embodiment disclosure of ERV2 may or may not also include
the combinations of ERV2 and ERO1, or ERO1 in its place.
[0228] In another embodiment, all seven of the above helper
proteins may or may not be simultaneously over-expressed in the
host cell. In that case, the following combinations may or may not
be chosen--
ERO1, ERV2, EUG1, MPD1, MPD2, EPS1 and PDI1.
[0229] Where the host cell is genetically modified to cause
simultaneous over-expression of one or two of the above defined
disulphide bond formation helper proteins, it may or may not be
preferred that the host cell is genetically modified to cause
simultaneous over-expression of at least three helper proteins and
the one or two other helper proteins may or may not be chaperones
or helper proteins involved in protein degradation, as discussed
above, and below, respectively.
[0230] Where one of the helper proteins is a protein disulphide
isomerase, such as a yeast and mammalian PDI, mammalian Erp59,
mammalian prolyl-4-hydroxylase B-subunit, yeast GSBP, yeast EUG1
and mammalian T3BP, then it may or may not be preferred, in one
embodiment, to avoid co-expression with KAR2 or an equivalent
thereof including hsp chaperone proteins such as other yeast Hsp70
proteins, BiP, SSA1-4, SSB1, SSC1 and SSD1 gene products and
eukaryotic hsp70 proteins such as HSP68, HSP72, HSP73, HSC70,
clathrin uncoating ATPase, IgG heavy chain binding protein (BiP),
glucose-regulated proteins 75, 78 and 80 (GRP75, GPR78 and GRP80)
and the like, particularly where these are the sole helper proteins
that are overexpressed in the host cell.
Proteins Involved in Protein Degradation
[0231] Proteins involved in protein degradation include DER1, DER3,
HRD3, UBC7 and DOA4. A detailed description of these proteins and
their genes is given separately below.
[0232] In one embodiment, one of the above proteins involved in
protein degradation may or may not be over-expressed in the host
cell. For example, DER1 may or may not be chosen, DER3 may or may
not be chosen, HRD3 may or may not be chosen, UBC7 may or may not
be chosen, or DOA4 may or may not be chosen.
[0233] In another embodiment, two of the above proteins involved in
protein degradation may or may not be simultaneously over-expressed
in the host cell. For example, one of the following combinations
may or may not be chosen--
DER1 and DER3; DER1 and HRD3; DER1 and UBC7; DER1 and DOA4; DER3
and HRD3; DER3 and UBC7; DER3 and DOA4; HRD3 and UBC7; HRD3 and
DOA4; or UBC7 and DOA4.
[0234] In another embodiment, three of the above proteins involved
in protein degradation may or may not be simultaneously
over-expressed in the host cell. For example, one of the following
combinations may or may not be chosen--
DER1, DER3 and HRD3; DER1, DER3 and UBC7; DER1, DER3 and DOA4;
DER1, HRD3 and UBC7; DER1, HRD3 and DOA4; DER1, UBC7 and DOA4;
DER3, HRD3 and UBC7; DER3, HRD3 and DOA4; DER3, UBC7 and DOA4; or
HRD3, UBC7 and DOA4.
[0235] In another embodiment, four of the above proteins involved
in protein degradation may or may not be simultaneously
over-expressed in the host cell. For example, one of the following
combinations may or may not be chosen--
DER1, DER3, HRD3 and UBC7; DER1, DER3, HRD3 and DOA4; DER1, DER3,
UBC7 and DOA4; DER1, HRD3, UBC7 and DOA4; or DER3, HRD3, UBC7 and
DOA4.
[0236] In another embodiment, all five of the above proteins
involved in protein degradation may or may not be simultaneously
over-expressed in the host cell. In that case, the following
combination is chosen--
DER1, DER3, HRD3, UBC7 and DOA4.
[0237] Where the host cell is genetically modified to cause
simultaneous over-expression of one or two of the above defined
protein degradation helper proteins, it may or may not be preferred
that the host cell is genetically modified to cause simultaneous
over-expression of at least three helper proteins in total and the
one or two other helper proteins may or may not be chaperones or
disulphide bond formation helper proteins, as discussed above.
HAC1 (Encoded by a Spliced or Unspliced Polynucleotide)
[0238] Valkonen et al. 2003 (Applied Environ. Micro., 69, 2065)
reported investigations into the possibility to obtain better
yields of secreted proteins. The authors found that the
manipulation of the unfolded-protein response (UPR) pathway
regulator, HAC1, affected the production of both native and foreign
proteins in the yeast Saccharomyces cerevisiae. For example, it is
reported that constitutive over-expression of HAC1 caused a 70%
increase in alpha-amylase secretion. WO 01/72783 also reports that.
HAC1 overexpression can be used to increase the secretion of a
heterologous protein in a eukaryotic cell by inducing an elevated
UPR, and PTC2 and IRE1 are also suggested for use in place of
HAC1.
[0239] Over-expression of HAC1 can be achieved, for example, by the
introduction of a recombinant polynucleotide that comprises the
endogenous. HAC1 gene coding sequence or a truncated intronless
HAC1 coding sequence (Valkonen et al. 2003, Applied Environ.
Micro., 69, 2065). A detailed description of this protein and its
gene is given separately below. The same techniques can be used to
over-express PTC2 or IRE1.
[0240] In one embodiment of the present invention, a host cell of
the present invention may or may not be genetically engineered to
cause over-expression HAC1, PTC2 or IRE1, such as by modification
of an endogenous gene encoding HAC1, PTC2 or IRE1, or by
transformation with a recombinant gene encoding HAC1, PTC2 or IRE1.
For example HAC1, PTC2 or IRE1 may or may not be simultaneously
over-expressed with any of the above-defined combinations of other
helper proteins.
[0241] In one embodiment, the host cell of the present invention is
not genetically engineered to cause HAC1 over-expression, such as
by modification of an endogenous HAC1 gene or transformation with a
recombinant HAC1 gene.
[0242] In another embodiment where the host cell is genetically
engineered to cause over-expression of HAC1, PTC2 or IRE1, the host
cell is additionally genetically modified by the introduction of at
least one recombinant gene encoding at least one other helper
protein, such as a DnaJ-like protein, an Hsp70 family protein
and/or SIL1 or by the modification of the sequence of an endogenous
gene encoding one or more other helper proteins at least one of a
DnaJ-like protein, an Hsp70 family protein (such as LHS1) and SIL1
to cause increased expression of the thus modified gene.
Other Combinations
[0243] In light of the above disclosure, the skilled person will
appreciate that the present invention also encompasses simultaneous
over-expression of any combination of helper proteins derived from
any of the above-defined groups.
[0244] For example, two helper proteins may or may not be
simultaneously over-expressed. Suitable combinations include any
one of the following combinations: JEM1 and LHS1; JEM1 and SCJ1;
JEM1 and KAR2; JEM1 and SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1
and SSA2; JEM1 and SSA3; JEM1 and SSA4; JEM1 and SSE1; JEM1 and
SSE2; JEM1 and SSB1; JEM1 and SSB2; JEM1 and ECM10; JEM1 and MDJ1;
JEM1 and MDJ2; JEM1 and ERO1; JEM1 and ERV2; JEM1 and EUG1; JEM1
and MPD1; JEM1 and MPD2; JEM1 and EPS1; JEM1 and PDI1; JEM1 and
DER1; JEM1 and DER3; JEM1 and HRD3; JEM1 and UBC7; JEM1 and DOA4;
JEM1 and HAC1; LHS1 and SCJ1; LHS1 and KAR2; LHS1 and SIL1 LHS1 and
FKB2; LHS1 and SSA1; LHS1 and SSA2; LHS1 and SSA3; LHS1 and SSA4;
LHS1 and SSE1; LHS1 and SSE2; LHS1 and SSB1; LHS1 and SSB2; LHS1
and ECM10; LHS1 and MDJ1; LHS1 and MDJ2; LHS1 and ERO1; LHS1 and
ERV2; LHS1 and EUG1; LHS1 and MPD1; LHS1 and MPD2; LHS1 and EPS1;
LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1 and HRD3; LHS1
and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and KAR2; SCJ1 and
SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2; SCJ1 and SSA3;
SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1 and SSB1; SCJ1
and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and MDJ2; SCJ1 and
ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSB2; KAR2 and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and
ERO1; KAR2 and ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2;
KAR2 and EPS1; KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2
and HRD3; KAR2 and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and
FKB2; SIL1 and SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4;
SIL1 and SSE1; SIL1 and SSE2; SIL1 and SSB1; SIL1 and SSB2; SIL1
and ECM10; SIL1 and MDJ1; SIL1 and MDJ2; SIL1 and ERO1; SIL1 and
ERV2; SIL1 and EUG1; SIL1 and MPD1; SIL1 and MPD2; SIL1 and EPS1;
SIL1 and PDI1; SIL1 and DER1; SIL1 and DER3; SIL1 and HRD3; SIL1
and UBC7; SIL1 and DOA4; SIL1 and HAC1; FKB2 and SSA1; FKB2 and
SSA2; FKB2 and SSA3; FKB2 and SSA4; FKB2 and SSE1; FKB2 and SSE2;
FKB2 and SSB1; FKB2 and SSB2; FKB2 and ECM10; FKB2 and MDJ1; FKB2
and MDJ2; FKB2 and ERO1; FKB2 and ERV2; FKB2 and EUG1; FKB2 and
MPD1; FKB2 and MPD2; FKB2 and EPS1; FKB2 and PDI1; FKB2 and DER1;
FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7; FKB2 and DOA4; FKB2
and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1 and SSA4; SSA1 and
SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and SSB2; SSA1 and ECM10;
SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1; SSA1 and ERV2; SSA1
and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1 and
PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and UBC7;
SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4; SSA2
and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2 and
ECM10; SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and ERV2;
SSA2 and EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2
and PDI1; SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and
UBC7; SSA2 and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1;
SSA3 and SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3
and MDJ1; SSA3 and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and
EUG1; SSA3 and MPD1; SSA3 and MPD2; SSA3 and EPS1; SSA3 and PDI1;
SSA3 and DER1; SSA3 and DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3
and DOA4; SSA3 and HAC1; SSA4 and SSE1; SSA4 and SSE2; SSA4 and
SSB1; SSA4 and SSB2; SSA4 and ECM10; SSA4 and MDJ1; SSA4 and MDJ2;
SSA4 and ERO1; SSA4 and ERV2; SSA4 and EUG1; SSA4 and MPD1; SSA4
and MPD2; SSA4 and EPS1; SSA4 and PDI1; SSA4 and DER1; SSA4 and
DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4 and DOA4; SSA4 and HAC1;
SSE1 and SSE2; SSE1 and SSB1; SSE1 and SSB2; SSE1 and ECM10; SSE1
and MDJ1; SSE1 and MDJ2; SSE1 and ERO1; SSE1 and ERV2; SSE1 and
EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1 and EPS1; SSE1 and PDI1;
SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and UBC7; SSE1
and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2; SSE2 and
ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2 and ERV2;
SSE2 and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2
and PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and
UBC7; SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10;
SSB1 and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1
and EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and
PDI1; SSB1 and DER1; SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7;
SSB1 and DOA4; SSB1 and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2
and MDJ2; SSB2 and ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and
MPD1; SSB2 and MPD2; SSB2 and EPS1; SSB2 and PDI1; SSB2 and DER1;
SSB2 and DER3; SSB2 and HRD3; SSB2 and UBC7; SSB2 and DOA4; SSB2
and HAC1; ECM10 and MDJ1; ECM10 and MDJ2; ECM10 and ERO1; ECM10 and
ERV2; ECM10 and EUG1; ECM10 and MPD1; ECM10 and MPD2; ECM10 and
EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and DER3; ECM10 and
HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and HAC1; MDJ1 and
MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1 and MPD1;
MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2
and DER3; MDJ2 and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and
HAC1; ERO1 and ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2;
ERO1 and EPS1; ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1
and HRD3; ERO1 and UBC7; ERO1 and DOA4; ERO1 and HAC1; ERV2 and
EUG1; ERV2 and MPD1; ERV2 and MPD2; ERV2 and EPS1; ERV2 and PDI1;
ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3; ERV2 and UBC7; ERV2
and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and
EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and
HRD3; MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1;
MPD2 and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2
and UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and
DER1; EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PIM and
UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and HRD3;
DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3; DER3
and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3 and
DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; DOA4 and
HAC1.
[0245] The skilled person will also appreciate that the present
invention encompasses simultaneous over-expression of at least
three helper proteins, and that the at least three helper proteins
may or may not be taken from any combination of helper proteins
derived from any of the above-defined groups.
[0246] For example, one of the following combinations of three
helper proteins may or may not be simultaneously over-expressed,
with or without the over-expression of one or more additional
helper proteins:
[0247] JEM1 in combination with any one of the following
combinations: LHS1 and SCJ1; LHS1 and KAR2; LHS1 and SIL1 LHS1 and
FKB2; LHS1 and SSA1; LHS1 and SSA2; LHS1 and SSA3; LHS1 and SSA4;
LHS1 and SSE1; LHS1 and SSE2; LHS1 and SSB1; LHS1 and SSB2; LHS1
and ECM10; LHS1 and MDJ1; LHS1 and MDJ2; LHS1 and ERO1; LHS1 and
ERV2; LHS1 and EUG1; LHS1 and MPD1; LHS1 and MPD2; LHS1 and EPS1;
LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1 and HRD3; LHS1
and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and KAR2; SCJ1 and
SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2; SCJ1 and SSA3;
SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1 and SSB1; SCJ1
and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and MDJ2; SCJ1 and
ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSB2; KAR2 and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and
ERO1; KAR2 and ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2;
KAR2 and EPS1; KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2
and HRD3; KAR2 and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and
FKB2; SIL1 and SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4;
SIL1 and SSE1; SIL1 and SSE2; SIL1 and SSB1; SIL1 and SSB2; SIL1
and ECM10; SIL1 and MDJ1; KU and MDJ2; SIL1 and ERO1; SIL1 and
ERV2; SIL1 and EUG1; SIL1 and MPD1; SIL1 and MPD2; SIL1 and EPS1;
SIL1 and PDI1; SIL1 and DER1; SIL1 and DER3; SIL1 and HRD3; SIL1
and UBC7; SIL1 and DOA4; SIL1 and HAC1; FKB2 and SSA1; FKB2 and
SSA2; FKB2 and SSA3; FKB2 and SSA4; FKB2 and SSE1; FKB2 and SSE2;
FKB2 and SSB1; FKB2 and SSB2; FKB2 and ECM10; FKB2 and MDJ1; FKB2
and MDJ2; FKB2 and ERO1; FKB2 and ERV2; FKB2 and EUG1; FKB2 and
MPD1, FKB2 and MPD2; FKB2 and EPS1; FKB2 and PDI1; FKB2 and DER1;
FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7; FKB2 and DOA4; FKB2
and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1 and SSA4; SSA1 and
SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and SSB2; SSA1 and ECM10;
SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1; SSA1 and ERV2; SSA1
and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1 and
PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and UBC7;
SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4; SSA2
and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2 and
ECM10; SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and ERV2;
SSA2 and EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2
and PDI1; SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and
UBC7; SSA2 and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1;
SSA3 and SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3
and MDJ1; SSA3 and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and
EUG1; SSA3 and MPD1; SSA3 and MPD2; SSA3 and EPS1; SSA3 and PDI1;
SSA3 and DER1; SSA3 and DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3
and DOA4; SSA3 and HAC1; SSA4 and SSE1; SSA4 and SSE2; SSA4 and
SSB1; SSA4 and SSB2; SSA4 and ECM10; SSA4 and MDJ1; SSA4 and MDJ2;
SSA4 and ERO1; SSA4 and ERV2; SSA4 and EUG1; SSA4 and MPD1; SSA4
and MPD2; SSA4 and EPS1; SSA4 and PDI1; SSA4 and DER1; SSA4 and
DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4 and DOA4; SSA4 and HAC1;
SSE1 and SSE2; SSE1 and SSB1; SSE1 and SSB2; SSE1 and ECM10; SSE1
and MDJ1; SSE1, and MDJ2; SSE1 and ERO1; SSE1 and ERV2; SSE1 and
EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1 and EPS1; SSE1 and PDI1;
SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and UBC7; SSE1
and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2; SSE2 and
ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2 and ERV2;
SSE2 and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2
and PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and
UBC7; SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10;
SSB1 and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1
and EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and
PDI1; SSB1 and DER1; SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7;
SSB1 and DOA4; SSB1 and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2
and MDJ2; SSB2 and ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and
MPD1; SSB2 and MPD2; SSB2 and EPS1; SSB2 and PDI1; SSB2 and DER1;
SSB2 and DER3; SSB2 and HRD3; SSB2 and UBC7; SSB2 and DOA4; SSB2
and HAC1; ECM10 and MDJ1; ECM10 and MDJ2; ECM10 and ERO1; ECM10 and
ERV2; ECM10 and EUG1; ECM10 and MPD1; ECM10 and MPD2; ECM10 and
EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and DER3; ECM10 and
HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and HAC1; MDJ1 and
MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1 and MPD1;
MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2
and DER3; MDJ2 and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and
HAC1; ERO1 and ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2;
ERO1 and EPS1; ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1
and HRD3; ERO1 and UBC7; ERO1 and DOA4; ERO1 and HAC1; ERV2 and
EUG1; ERV2 and MPD1; ERV2 and MPD2; ERV2 and EPS1; ERV2 and PDI1;
ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3; ERV2 and UBC7; ERV2
and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and
EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and
HRD3; MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1;
MPD2 and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2
and UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and
DER1; EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0248] LHS1 in combination with any one of the following
combinations: JEM1 and SCJ1; JEM1 and KAR2; JEM1 and SIL1; JEM1 and
FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3; JEM1 and SSA4;
JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1 and SSB2; JEM1
and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and ERO1; JEM1 and
ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; SCJ1 and KAR2; SCJ1 and
SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2; SCJ1 and SSA3;
SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1 and SSB1; SCJ1
and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and MDJ2; SCJ1 and
ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSB2; KAR2 and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and
ERO1; KAR2 and ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2;
KAR2 and EPS1; KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2
and HRD3; KAR2 and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and
FKB2; SIL1 and SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4;
SIL1 and SSE1; SIL1 and SSE2; SIL1 and SSB1; SIL1 and SSB2; SIL1
and ECM10; SIL1 and MDJ1; SIL1 and MDJ2; SIL1 and ERO1; SIL1 and
ERV2; SIL1 and EUG1; SIL1 and MPD1; SIL1 and MPD2; SIL1 and EPS1;
SIL1 and PDI1; SIL1, and DER1; SIL1 and DER3; SIL1 and HRD3; SIL1
and UBC7; SIL1 and DOA4; SIL1 and HAC1; FKB2 and SSA1; FKB2 and
SSA2; FKB2 and SSA3; FKB2 and SSA4; FKB2 and SSE1; FKB2 and SSE2;
FKB2 and SSB1; FKB2 and SSB2; FKB2 and ECM10; FKB2 and MDJ1; FKB2
and MDJ2; FKB2 and ERO1; FKB2 and ERV2; FKB2 and EUG1; FKB2 and
MPD1; FKB2 and MPD2; FKB2 and EPS1; FKB2 and PDI1; FKB2 and DER1;
FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7; FKB2 and DOA4; FKB2
and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1 and SSA4; SSA1 and
SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and SSB2; SSA1 and ECM10;
SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1; SSA1 and ERV2; SSA1
and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1 and
PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and UBC7;
SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4; SSA2
and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2 and
ECM10; SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and ERV2;
SSA2 and EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2
and PDI1; SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and
UBC7; SSA2 and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1;
SSA3 and SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3
and MDJ1; SSA3 and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and
EUG1; SSA3 and MPD1; SSA3 and MPD2; SSA3 and EPS1; SSA3 and PDI1;
SSA3 and DER1; SSA3 and DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3
and DOA4; SSA3 and HAC1; SSA4 and SSE1; SSA4 and SSE2; SSA4 and
SSB1; SSA4 and SSB2; SSA4 and ECM10; SSA4 and MDJ1; SSA4 and MDJ2;
SSA4 and ERO1; SSA4 and ERV2; SSA4 and EUG1; SSA4 and MPD1; SSA4
and MPD2; SSA4 and EPS1; SSA4 and PDI1; SSA4 and DER1; SSA4 and
DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4 and DOA4; SSA4 and HAC1;
SSE1 and SSE2; SSE1 and SSB1; SSE1 and SSB2; SSE1 and ECM10; SSE1
and MDJ1; SSE1 and MDJ2; SSE1 and ERO1; SSE1 and ERV2; SSE1 and
EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1 and EPS1; SSE1 and PDI1;
SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and UBC7; SSE1
and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2; SSE2 and
ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2 and ERV2;
SSE2 and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2
and PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and
UBC7; SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10;
SSB1 and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1
and EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and
PDI1; SSB1 and DER1; SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7;
SSB1 and DOA4; SSB1 and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2
and MDJ2; SSB2 and ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and
MPD1; SSB2 and MPD2; SSB2 and EPS1; SSB2 and PDI1; SSB2 and DER1;
SSB2 and DER3; SSB2 and HRD3; SSB2 and UBC7; SSB2 and DOA4; SSB2
and HAC1; ECM10 and MDJ1; ECM10 and MDJ2; ECM10 and ERO1; ECM10 and
ERV2; ECM10 and EUG1; ECM10 and MPD1; ECM10 and MPD2; ECM10 and
EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and DER3; ECM10 and
HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and HAC1; MDJ1 and
MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1 and MPD1;
MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2
and DER3; MDJ2 and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and
HAC1; ERO1 and ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2;
ERO1 and EPS1; ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1
and HRD3; ERO1 and UBC7; ERO1 and DOA4; ERO1 and HAC1; ERV2 and
EUG1; ERV2 and MPD1; ERV2 and MPD2; ERV2 and EPS1; ERV2 and PDI1;
ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3; ERV2 and UBC7; ERV2
and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and
EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and
HRD3; MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1;
MPD2 and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2
and UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and
DER1; EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0249] SCJ1 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and KAR2; JEM1 and SIL1; JEM1 and
FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3; JEM1 and SSA4;
JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1 and SSB2; JEM1
and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and ERO1; JEM1 and
ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and KAR2; LHS1 and
SIL1; LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2; LHS1 and SSA3;
LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1 and SSB1; LHS1
and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and MDJ2; LHS1 and
ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1; LHS1 and MPD2;
LHS1 and EPS1; LHS1, and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSB2; KAR2 and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and
ERO1; KAR2 and ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2;
KAR2 and EPS1; KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2
and HRD3; KAR2 and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and
FKB2; SIL1 and SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4;
SIL1 and SSE1; SIL1 and SSE2; SIL1 and SSB1; SIL1 and SSB2; SIL1
and ECM10; SIL1 and MDJ1; SIL1 and MDJ2; SIL1 and ERO1; SIL1 and
ERV2; SIL1 and EUG1; SIL1 and MPD1; SIL1 and MPD2; SIL1 and EPS1;
SIL1 and PDI1; SIL1 and DER1; SIL1 and DER3; HU and HRD3; SIL1 and
UBC7; SIL1 and DOA4; SIL1 and HAC1; FKB2 and SSA1; FKB2 and SSA2;
FKB2 and SSA3; FKB2 and SSA4; FKB2 and SSE1; FKB2 and SSE2; FKB2
and SSB1; FKB2 and SSB2; FKB2 and ECM10; FKB2 and MDJ1; FKB2 and
MDJ2; FKB2 and ERO1; FKB2 and ERV2; FKB2 and EUG1; FKB2 and MPD1;
FKB2 and MPD2; FKB2 and EPS1; FKB2 and PDI1; FKB2 and DER1; FKB2
and DER3; FKB2 and HRD3; FKB2 and UBC7; FKB2 and DOA4; FKB2 and
HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1 and SSA4; SSA1 and SSE1;
SSA1 and SSE2; SSA1 and SSB1; SSA1 and SSB2; SSA1 and ECM10; SSA1
and MDJ1; SSA1 and MDJ2; SSA1 and ERO1; SSA1 and ERV2; SSA1 and
EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1 and PDI1;
SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and UBC7; SSA1
and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4; SSA2 and
SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2 and ECM10;
SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and ERV2; SSA2
and EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2 and
PDI1; SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and UBC7;
SSA2 and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3
and SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3 and
MDJ1; SSA3 and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and EUG1;
SSA3 and MPD1; SSA3 and MPD2; SSA3 and EPS1; SSA3 and PDI1; SSA3
and DER1; SSA3 and DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3 and
DOA4; SSA3 and HAC1; SSA4 and SSE1; SSA4 and SSE2; SSA4 and SSB1;
SSA4 and SSB2; SSA4 and ECM10; SSA4 and MDJ1; SSA4 and MDJ2; SSA4
and ERO1; SSA4 and ERV2; SSA4 and EUG1; SSA4 and MPD1; SSA4 and
MPD2; SSA4 and EPS1; SSA4 and PDI1; SSA4 and DER1; SSA4 and DER3;
SSA4 and HRD3; SSA4 and UBC7; SSA4 and DOA4; SSA4 and HAC1; SSE1
and SSE2; SSE1 and SSB1; SSE1 and SSB2; SSE1 and ECM10; SSE1 and
MDJ1; SSE1 and MDJ2; SSE1 and ERO1; SSE1 and ERV2; SSE1 and EUG1;
SSE1 and MPD1; SSE1 and MPD2; SSE1 and EPS1; SSE1 and PDI1; SSE1
and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and UBC7; SSE1 and
DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2; SSE2 and ECM10;
SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2 and ERV2; SSE2
and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2 and
PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and UBC7;
SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10; SSB1
and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1 and
EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and PDI1;
SSB1 and DER1; SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7; SSB1
and DOA4; SSB1 and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2 and
MDJ2; SSB2 and ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and MPD1;
SSB2 and MPD2; SSB2 and EPS1; SSB2 and PDI1; SSB2 and DER1; SSB2
and DER3; SSB2 and HRD3; SSB2 and UBC7; SSB2 and DOA4; SSB2 and
HAC1; ECM10 and MDJ1; ECM10 and MDJ2; ECM10 and ERO1; ECM10 and
ERV2; ECM10 and EUG1; ECM10 and MPD1; ECM10 and MPD2; ECM10 and
EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and DER3; ECM10 and
HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and HAC1; MDJ1 and
MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1 and MPD1;
MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2
and DER3; MDJ2 and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and
HAC1; ERO1 and ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2;
ERO1 and EPS1; ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1
and HRD3; ERO1 and UBC7; ERO1 and DOA4; ERO1 and HAC1; ERV2 and
EUG1; ERV2 and MPD1; ERV2 and MPD2; ERV2 and EPS1; ERV2 and PDI1;
ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3; ERV2 and UBC7; ERV2
and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and
EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and
HRD3; MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1;
MPD2 and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2
and UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and
DER1; EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0250] KAR2 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and SIL1; JEM1 and
FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3; JEM1 and SSA4;
JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1 and SSB2; JEM1
and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and ERO1; JEM1 and
ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
SIL1 LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2; LHS1 and SSA3;
LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1 and SSB1; LHS1
and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and MDJ2; LHS1 and
ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1; LHS1 and MPD2;
LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2; SCJ1 and SSA3;
SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1 and SSB1; SCJ1
and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and MDJ2; SCJ1 and
ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; SIL1 and
FKB2; SIL1 and SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4;
SIL1 and SSE1; SIL1 and SSE2; SIL1 and SSB1; SIL1 and SSB2; SIL1
and ECM10; SIL1 and MDJ1; SIL1 and MDJ2; SIL1 and ERO1; SIL1 and
ERV2; SIL1 and EUG1; SIL1 and MPD1; SIL1 and MPD2; SIL1 and EPS1;
SIL1 and PDI1; SIL1 and DER1; SIL1 and DER3; SIL1 and HRD3; SEM and
UBC7; SIL1 and DOA4; SIL1 and HAC1; FKB2 and SSA1; FKB2 and SSA2;
FKB2 and SSA3; FKB2 and SSA4; FKB2 and SSE1; FKB2 and SSE2; FKB2
and SSB1; FKB2 and SSB2; FKB2 and ECM10; FKB2 and MDJ1; FKB2 and
MDJ2; FKB2 and ERO1; FKB2 and ERV2; FKB2 and EUG1; FKB2 and MPD1;
FKB2 and MPD2; FKB2 and EPS1; FKB2 and PDI1; FKB2 and DER1; FKB2
and DER3; FKB2 and HRD3; FKB2 and UBC7; FKB2 and DOA4; FKB2 and
HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1 and SSA4; SSA1 and SSE1;
SSA1 and SSE2; SSA1 and SSB1; SSA1 and SSB2; SSA1 and ECM10; SSA1
and MDJ1; SSA1 and MDJ2; SSA1 and ERO1; SSA1 and ERV2; SSA1 and
EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1 and PDI1;
SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and UBC7; SSA1
and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4; SSA2 and
SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2 and ECM10;
SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and ERV2; SSA2
and EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2 and
PDI1; SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and UBC7;
SSA2 and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3
and SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3 and
MDJ1; SSA3 and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and EUG1;
SSA3 and MPD1; SSA3 and MPD2; SSA3 and EPS1; SSA3 and PDI1; SSA3
and DER1; SSA3 and DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3 and
DOA4; SSA3 and HAC1; SSA4 and SSE1; SSA4 and SSE2; SSA4 and SSB1;
SSA4 and SSB2; SSA4 and ECM10; SSA4 and MDJ1; SSA4 and MDJ2; SSA4
and ERO1; SSA4 and ERV2; SSA4 and EUG1; SSA4 and MPD1; SSA4 and
MPD2; SSA4 and EPS1; SSA4 and PDI1; SSA4 and DER1; SSA4 and DER3;
SSA4 and HRD3; SSA4 and UBC7; SSA4 and DOA4; SSA4 and HAC1; SSE1
and SSE2; SSE1 and SSB1; SSE1 and SSB2; SSE1 and ECM10; SSE1 and
MDJ1; SSE1 and MDJ2; SSE1 and ERO1; SSE1 and ERV2; SSE1 and EUG1;
SSE1 and MPD1; SSE1 and MPD2; SSE1 and EPS1; SSE1 and PDI1; SSE1
and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and UBC7; SSE1 and
DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2; SSE2 and ECM10;
SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2 and ERV2; SSE2
and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2 and
PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and UBC7;
SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10; SSB1
and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1 and
EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and PDI1;
SSB1 and DER1; SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7; SSB1
and DOA4; SSB1 and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2 and
MDJ2; SSB2 and ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and MPD1;
SSB2 and MPD2; SSB2 and EPS1; SSB2 and PDI1; SSB2 and DER1; SSB2
and DER3; SSB2 and HRD3; SSB2 and UBC7; SSB2 and DOA4; SSB2 and
HAC1; ECM10 and MDJ1; ECM10 and MDJ2; ECM10 and ERO1; ECM10 and
ERV2; ECM10 and EUG1; ECM10 and MPD1; ECM10 and MPD2; ECM10 and
EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and DER3; ECM10 and
HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and HAC1; MDJ1 and
MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1 and MPD1;
MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2
and DER3; MDJ2 and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and
HAC1; ERO1 and ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2;
ERO1 and EPS1; ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1
and HRD3; ERO1 and UBC7; ERO1 and DOA4; ERO1 and HAC1; ERV2 and
EUG1; ERV2 and MPD1; ERV2 and MPD2; ERV2 and EPS1; ERV2 and PDI1;
ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3; ERV2 and UBC7; ERV2
and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and
EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and
HRD3; MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1;
MPD2 and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2
and UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and
DER1; EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0251] SIL1 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3; JEM1 and SSA4;
JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1 and SSB2; JEM1
and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and ERO1; JEM1 and
ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2; LHS1 and SSA3;
LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1 and SSB1; LHS1
and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and MDJ2; LHS1 and
ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1; LHS1 and MPD2;
LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2; SCJ1 and SSA3;
SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1 and SSB1; SCJ1
and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and MDJ2; SCJ1 and
ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3; KAR2 and SSA4;
KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2 and SSB2; KAR2
and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and ERO1; KAR2 and
ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2; KAR2 and EPS1;
KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; FKB2 and SSA1; FKB2 and
SSA2; FKB2 and SSA3; FKB2 and SSA4; FKB2 and SSE1; FKB2 and SSE2;
FKB2 and SSB1; FKB2 and SSB2; FKB2 and ECM10; FKB2 and MDJ1; FKB2
and MDJ2; FKB2 and ERO1; FKB2 and ERV2; FKB2 and EUG1; FKB2 and
MPD1; FKB2 and MPD2; FKB2 and EPS1; FKB2 and PDI1; FKB2 and DER1;
FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7; FKB2 and DOA4; FKB2
and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1 and SSA4; SSA1 and
SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and SSB2; SSA1 and ECM10;
SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1; SSA1 and ERV2; SSA1
and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1 and
PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and UBC7;
SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4; SSA2
and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2 and
ECM10; SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and ERV2;
SSA2 and EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2
and PDI1; SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and
UBC7; SSA2 and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1;
SSA3 and SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3
and MDJ1; SSA3 and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and
EUG1; SSA3 and MPD1; SSA3 and MPD2; SSA3 and EPS1; SSA3 and PDI1;
SSA3 and DER1; SSA3 and DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3
and DOA4; SSA3 and HAC1; SSA4 and SSE1; SSA4 and SSE2; SSA4 and
SSB1; SSA4 and SSB2; SSA4 and ECM10; SSA4 and MDJ1; SSA4 and MDJ2;
SSA4 and ERO1; SSA4 and ERV2; SSA4 and EUG1; SSA4 and MPD1; SSA4
and MPD2; SSA4 and EPS1; SSA4 and PDI1; SSA4 and DER1; SSA4 and
DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4 and DOA4; SSA4 and HAC1;
SSE1 and SSE2; SSE1 and SSB1; SSE1 and SSB2; SSE1 and ECM10; SSE1
and MDJ1; SSE1 and MDJ2; SSE1 and ERO1; SSE1 and ERV2; SSE1 and
EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1 and EPS1; SSE1 and PDI1;
SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and UBC7; SSE1
and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2; SSE2 and
ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2 and ERV2;
SSE2 and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2
and PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and
UBC7; SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10;
SSB1 and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1
and EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and
PDI1; SSB1 and DER1; SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7;
SSB1 and DOA4; SSB1 and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2
and MDJ2; SSB2 and ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and
MPD1; SSB2 and MPD2; SSB2 and EPS1; SSB2 and PDI1; SSB2 and DER1;
SSB2 and DER3; SSB2 and HRD3; SSB2 and UBC7; SSB2 and DOA4; SSB2
and HAC1; ECM10 and MDJ1; ECM10 and MDJ2; ECM10 and ERO1; ECM10 and
ERV2; ECM10 and EUG1; ECM10 and MPD1; ECM10 and MPD2; ECM10 and
EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and DER3; ECM10 and
HRD3; ECM10, and UBC7; ECM10 and DOA4; ECM10 and HAC1; MDJ1 and
MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1 and MPD1;
MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2
and DER3; MDJ2 and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and
HAC1; ERO1 and ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2;
ERO1 and EPS1; ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1
and HRD3; ERO1 and UBC7; ERO1 and DOA4; ERO1 and HAC1; ERV2 and
EUG1; ERV2 and MPD1; ERV2 and MPD2; ERV2 and EPS1; ERV2 and PDI1;
ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3; ERV2 and UBC7; ERV2
and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and
EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and
HRD3; MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1;
MPD2 and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2
and UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and
DER1; EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0252] FKB2 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3; JEM1 and SSA4;
JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1 and SSB2; JEM1
and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and ERO1; JEM1 and
ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1 LHS1 and SSA1; LHS1 and SSA2; LHS1 and SSA3;
LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1 and SSB1; LHS1
and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and MDJ2; LHS1 and
ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1; LHS1 and MPD2;
LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and SSA1; SCJ1 and SSA2; SCJ1 and SSA3;
SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1 and SSB1; SCJ1
and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and MDJ2; SCJ1 and
ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3; KAR2 and SSA4;
KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2 and SSB2; KAR2
and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and ERO1; KAR2 and
ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2; KAR2 and EPS1;
KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and SSA1; SILT and
SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1; SIL1 and SSE2;
SIL1 and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1 and MDJ1; SIL1
and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and EUG1; SIL1 and
MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1; SIL1 and DER1;
SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1
and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1 and SSA4; SSA1 and
SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and SSB2; SSA1 and ECM10;
SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1; SSA1 and ERV2; SSA1
and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1 and
PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and UBC7;
SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4; SSA2
and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2 and
ECM10; SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and ERV2;
SSA2 and EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2
and PDI1; SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and
UBC7; SSA2 and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1;
SSA3 and SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3
and MDJ1; SSA3 and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and
EUG1; SSA3 and MPD1; SSA3 and MPD2; SSA3 and EPS1; SSA3 and PDI1;
SSA3 and DER1; SSA3 and DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3
and DOA4; SSA3 and HAC1; SSA4 and SSE1; SSA4 and SSE2; SSA4 and
SSB1; SSA4 and SSB2; SSA4 and ECM10; SSA4 and MDJ1; SSA4 and MDJ2;
SSA4 and ERO1; SSA4 and ERV2; SSA4 and EUG1; SSA4 and MPD1; SSA4
and MPD2; SSA4 and EPS1; SSA4 and PDI1; SSA4 and DER1; SSA4 and
DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4 and DOA4; SSA4 and HAC1;
SSE1 and SSE2; SSE1 and SSB1; SSE1 and SSB2; SSE1 and ECM10; SSE1
and MDJ1; SSE1 and MDJ2; SSE1 and ERO1; SSE1 and ERV2; SSE1 and
EUG1; SSE1 and MIDI; SSE1 and MPD2; SSE1 and EPS1; SSE1 and PDI1;
SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and UBC7; SSE1
and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2; SSE2 and
ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2 and ERV2;
SSE2 and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2
and PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and
UBC7; SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10;
SSB1 and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1
and EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and
PDI1; SSB1 and DER1; SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7;
SSB1 and DOA4; SSB1 and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2
and MDJ2; SSB2 and ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and
MPD1; SSB2 and MPD2; SSB2 and EPS1; SSB2 and PDI1; SSB2 and DER1;
SSB2 and DER3; SSB2 and HRD3; SSB2 and UBC7; SSB2 and DOA4; SSB2
and HAC1; ECM10 and MDJ1; ECM10 and MDJ2; ECM10 and ERO1; ECM10 and
ERV2; ECM10 and EUG1; ECM10 and MPD1; ECM10 and MPD2; ECM10 and
EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and DER3; ECM10 and
HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and HAC1; MDJ1 and
MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1 and MPD1;
MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2- and DER1; MDJ2
and DER3; MDJ2 and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and
HAC1; ERO1 and ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2;
ERO1 and EPS1; ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1
and HRD3; ERO1 and UBC7; ERO1 and DOA4; ERO1 and HAC1; ERV2 and
EUG1; ERV2 and MPD1; ERV2 and MPD2; ERV2 and EPS1; ERV2 and PDI1;
ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3; ERV2 and UBC7; ERV2
and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and
EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and
HRD3; MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1;
MPD2 and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2
and UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and
DER1; EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0253] SSA1 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA2; JEM1 and SSA3; JEM1 and SSA4;
JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1 and SSB2; JEM1
and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and ERO1; JEM1 and
ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1; LHS1 and FKB2; LHS1 and SSA2; LHS1 and SSA3;
LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1 and SSB1; LHS1
and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and MDJ2; LHS1 and
ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1; LHS1 and MPD2;
LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA2; SCJ1 and SSA3;
SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1 and SSB1; SCJ1
and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and MDJ2; SCJ1 and
ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA2; KAR2 and SSA3; KAR2 and SSA4;
KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2 and SSB2; KAR2
and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and ERO1; KAR2 and
ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2; KAR2 and EPS1;
KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA2; SIL1 and SSA3; SEM and SSA4; SIL1 and SSE1; SIL1 and SSE2;
SIL1 and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1 and MDJ1; SIL1
and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and EUG1; SIL1 and
MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1; SIL1 and DER1;
SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1
and HAC1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and SSA4; FKB2 and
SSE1; FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2; FKB2 and ECM10;
FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2 and ERV2; FKB2
and EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and EPS1; FKB2 and
PDI1; FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7;
FKB2 and DOA4; FKB2 and HAC1; SSA2 and SSA3; SSA2 and SSA4; SSA2
and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2 and
ECM10; SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and ERV2;
SSA2 and EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2
and PDI1; SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and
UBC7; SSA2 and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1;
SSA3 and SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3
and MDJ1; SSA3 and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and
EUG1; SSA3 and MPD1; SSA3 and MPD2; SSA3 and EPS1; SSA3 and PDI1;
SSA3 and DER1; SSA3 and DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3
DOA4; SSA3 and HAC1; SSA4 and SSE1; SSA4 acid SSE2; SSA4 and SSB1;
SSA4 and SSB2; SSA4 and ECM10; SSA4 and MDJ1; SSA4 and MDJ2; SSA4
and ERO1; SSA4 and ERV2; SSA4 and EUG1; SSA4 and MPD1; SSA4 and
MPD2; SSA4 and EPS1; SSA4 and PDI1; SSA4 and DER1; SSA4 and DER3;
SSA4 and HRD3; SSA4 and UBC7; SSA4 and DOA4; SSA4 and HAC1; SSE1
and SSE2; SSE1 and SSB1; SSE1 and SSB2; SSE1 and ECM10; SSE1 and
MDJ1; SSE1 and MDJ2; SSE1 and ERO1; SSE1 and ERV2; SSE1 and EUG1;
SSE1 and MPD1; SSE1 and MPD2; SSE1 and EPS1; SSE1 and PDI1; SSE1
and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and UBC7; SSE1 and
DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2; SSE2 and ECM10;
SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2 and ERV2; SSE2
and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2 and
PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and UBC7;
SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10; SSB1
and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1 and
EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and PDI1;
SSB1 and DER1; SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7; SSB1
and DOA4; SSB1 and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2 and
MDJ2; SSB2 and ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and MPD1;
SSB2 and MPD2; SSB2 and EPS1; SSB2 and PDI1; SSB2 and DER1; SSB2
and DER3; SSB2 and HRD3; SSB2 and UBC7; SSB2 and DOA4; SSB2 and
HAC1; ECM10 and MDJ1; ECM10 and MDJ2; ECM10 and ERO1; ECM10 and
ERV2; ECM10 and EUG1; ECM10 and MPD1; ECM10 and MPD2; ECM10 and
EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and DER3; ECM10 and
HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and HAC1; MDJ1 and
MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1 and MPD1;
MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2
and DER3; MDJ2 and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and
HAC1; ERO1 and ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2;
ERO1 and EPS1; ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1
and HRD3; ERO1 and UBC7; ERO1 and DOA4; ERO1 and HAC1; ERV2 and
EUG1; ERV2 and MPD1; ERV2 and MPD2; ERV2 and EPS1; ERV2 and PDI1;
ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3; ERV2 and UBC7; ERV2
and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and
EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and
HRD3; MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1;
MPD2 and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2
and UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and
DER1; EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0254] SSA2 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA3; JEM1 and SSA4;
JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1 and SSB2; JEM1
and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and ERO1; JEM1 and
ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1 LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA3;
LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1 and SSB1; LHS1
and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and MDJ2; LHS1 and
ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1; LHS1 and MPD2;
LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA3;
SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1 and SSB1; SCJ1
and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and MDJ2; SCJ1 and
ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA3; KAR2 and SSA4;
KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2 and SSB2; KAR2
and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and ERO1; KAR2 and
ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2; KAR2 and EPS1;
KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1; SIL1 and SSE2;
Sad and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1 and MDJ1; SIL1
and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and EUG1; SIL1 and
MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1; SIL1 and DER1;
SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1
and HAC1; FKB2 and SSA1; FKB2 and SSA3; FKB2 and SSA4; FKB2 and
SSE1; FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2; FKB2 and ECM10;
FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2 and ERV2; FKB2
and EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and EPS1; FKB2 and
PDI1; FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7;
FKB2 and DOA4; FKB2 and HAC1; SSA1 and SSA3; SSA1 and SSA4; SSA1
and SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and SSB2; SSA1 and
ECM10; SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1; SSA1 and ERV2;
SSA1 and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1
and PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and
UBC7; SSA1 and DOA4; SSA1 and HAC1; SSA3 and SSA4; SSA3 and SSE1;
SSA3 and SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3
and MDJ1; SSA3 and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and
EUG1; SSA3 and MPD1; SSA3 and MPD2; SSA3 and EPS1; SSA3 and PDI1;
SSA3 and DER1; SSA3 and DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3
and DOA4; SSA3 and HAC1; SSA4 and SSE1; SSA4 and SSE2; SSA4 and
SSB1; SSA4 and SSB2; SSA4 and ECM10; SSA4 and MDJ1; SSA4 and MDJ2;
SSA4 and ERO1; SSA4 and ERV2; SSA4 and EUG1; SSA4 and MPD1; SSA4
and MPD2; SSA4 and EPS1; SSA4 and PDI1; SSA4 and DER1; SSA4 and
DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4 and DOA4; SSA4 and HAC1;
SSE1 and SSE2; SSE1 and SSB1; SSE1 and SSB2; SSE1 and ECM10; SSE1
and MDJ1; SSE1 and MDJ2; SSE1 and ERO1; SSE1 and ERV2; SSE1 and
EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1 and EPS1; SSE1 and PDI1;
SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and UBC7; SSE1
and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2; SSE2 and
ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2 and ERV2;
SSE2 and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2
and PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and
UBC7; SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10;
SSB1 and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1
and EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and
PDI1; SSB1 and DER1; SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7;
SSB1 and DOA4; SSB1 and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2
and MDJ2; SSB2 and ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and
MPD1; SSB2 and MPD2; SSB2 and EPS1; SSB2 and PDI1; SSB2 and DER1;
SSB2 and DER3; SSB2 and HRD3; SSB2 and UBC7; SSB2 and DOA4; SSB2
and HAC1; ECM10 and MDJ1; ECM10 and MDJ2; ECM10 and ERO1; ECM10 and
ERV2; ECM10 and EUG1; ECM10 and MPD1; ECM10 and MPD2; ECM10 and
EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and DER3; ECM10 and
HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and HAC1; MDJ1 and
MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1 and MPD1;
MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2
and DER3; MDJ2 and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and
HAC1; ERO1 and ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2;
ERO1 and EPS1; ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1
and HRD3; ERO1 and UBC7; ERO1 and DOA4; ERO1 and HAC1; ERV2 and
EUG1; ERV2 and MPD1; ERV2 and MPD2; ERV2 and EPS1; ERV2 and PDI1;
ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3; ERV2 and UBC7; ERV2
and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and
EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and
HRD3; MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1;
MPD2 and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2
and UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and
DER1; EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0255] SSA3 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA4;
JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1 and SSB2; JEM1
and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and ERO1; JEM1 and
ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1; LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1 and SSB1; LHS1
and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and MDJ2; LHS1 and
ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1; LHS1 and MPD2;
LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCH and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1 and SSB1; SCJ1
and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and MDJ2; SCJ1 and
ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCH and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1 and
HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and SIL1;
KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA4; KAR2
and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2 and SSB2; KAR2 and
ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and ERO1; KAR2 and ERV2;
KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2; KAR2 and EPS1; KAR2
and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2 and
UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and SSA1;
SIL1 and SSA2; SIL1 and SSA4; SIL1 and SSE1; SIL1 and SSE2; SIL1
and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1 and MDJ1; SIL1 and
MDJ2; SIL1 and ERO1; SILT and ERV2; SIL1 and EUG1; SIL1 and MPD1;
SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1; SIL1 and DER1; SIL1
and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1 and
HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA4; FKB2 and SSE1;
FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2; FKB2 and ECM10; FKB2
and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2 and ERV2; FKB2 and
EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and EPS1; FKB2 and PDI1;
FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7; FKB2
and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA4; SSA1 and
SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and SSB2; SSA1 and ECM10;
SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1; SSA1 and ERV2; SSA1
and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1 and
PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and UBC7;
SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA4; SSA2 and SSE1; SSA2
and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2 and ECM10; SSA2 and
MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and ERV2; SSA2 and EUG1;
SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2 and PDI1; SSA2
and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and UBC7; SSA2 and
DOA4; SSA2 and HAC1; SSA4 and SSE1; SSA4 and SSE2; SSA4 and SSB1;
SSA4 and SSB2; SSA4 and ECM10; SSA4 and MDJ1; SSA4 and MDJ2; SSA4
and ERO1; SSA4 and ERV2; SSA4 and EUG1; SSA4 and MPD1; SSA4 and
MPD2; SSA4 and EPS1; SSA4 and PDI1; SSA4 and DER1; SSA4 and DER3;
SSA4 and HRD3; SSA4 and UBC7; SSA4 and DOA4; SSA4 and HAC1; SSE1
and SSE2; SSE1 and SSB1; SSE1 and SSB2; SSE1 and ECM10; SSE1 and
MDJ1; SSE1 and MDJ2; SSE1 and ERO1; SSE1 and ERV2; SSE1 and EUG1;
SSE1 and MPD1; SSE1 and MPD2; SSE1 and EPS1; SSE1 and PDI1; SSE1
and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and UBC7; SSE1 and
DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2; SSE2 and ECM10;
SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2 and ERV2; SSE2
and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2 and
PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and UBC7;
SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10; SSB1
and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1 and
EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and PDI1;
SSB1 and DER1; SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7; SSB1
and DOA4; SSB1 and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2 and
MDJ2; SSB2 and ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and MPD1;
SSB2 and MPD2; SSB2 and EPS1; SSB2 and PDI1; SSB2 and DER1; SSB2
and DER3; SSB2 and HRD3; SSB2 and UBC7; SSB2 and DOA4; SSB2 and
HAC1; ECM10 and MDJ1; ECM10 and MDJ2; ECM10 and ERO1; ECM10 and
ERV2; ECM10 and EUG1; ECM10 and MPD1; ECM10 and MPD2; ECM10 and
EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and DER3; ECM10 and
HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and HAC1; MDJ1 and
MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1 and MPD1;
MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2
and DER3; MDJ2 and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and
HAC1; ERO1 and ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2;
ERO1 and EPS1; ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1
and HRD3; ERO1 and UBC7; ERO1 and DOA4; ERO1 and HAC1; ERV2 and
EUG1; ERV2 and MPD1; ERV2 and MPD2; ERV2 and EPS1; ERV2 and PDI1;
ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3; ERV2 and UBC7; ERV2
and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and
EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and
HRD3; MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1;
MPD2 and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2
and UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and
DER1; EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0256] SSA4 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1 and SSB2; JEM1
and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and ERO1; JEM1 and
ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1; LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSE1; LHS1 and SSE2; LHS1 and SSB1; LHS1
and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and MDJ2; LHS1 and
ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1; LHS1 and MPD2;
LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1 and SSB1; SCJ1
and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and MDJ2; SCJ1 and
ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2 and SSB2; KAR2
and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and ERO1; KAR2 and
ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2; KAR2 and EPS1;
KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSE1; SIL1 and SSE2;
SIL1 and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1 and MDJ1; SIL1
and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and EUG1; SIL1 and
MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1; SIL1 and DER1;
SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; KU and DOA4; SIL1 and
HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and SSE1;
FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2; FKB2 and ECM10; FKB2
and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2 and ERV2; FKB2 and
EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and EPS1; FKB2 and PDI1;
FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7; FKB2
and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1 and
SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and SSB2; SSA1 and ECM10;
SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1; SSA1 and ERV2; SSA1
and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1 and
PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and UBC7;
SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSE1; SSA2
and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2 and ECM10; SSA2 and
MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and ERV2; SSA2 and EUG1;
SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2 and PDI1; SSA2
and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and UBC7; SSA2 and
DOA4; SSA2 and HAC1; SSA3 and SSE1; SSA3 and SSE2; SSA3 and SSB1;
SSA3 and SSB2; SSA3 and ECM10; SSA3 and MDJ1; SSA3 and MDJ2; SSA3
and ERO1; SSA3 and ERV2; SSA3 and EUG1; SSA3 and MPD1; SSA3 and
MPD2; SSA3 and EPS1; SSA3 and PDI1; SSA3 and DER1; SSA3 and DER3;
SSA3 and HRD3; SSA3 and UBC7; SSA3 and DOA4; SSA3 and HAC1; SSE1
and SSE2; SSE1 and SSB1; SSE1 and SSB2; SSE1 and ECM10; SSE1 and
MDJ1; SSE1 and MDJ2; SSE1 and ERO1; SSE1 and ERV2; SSE1 and EUG1;
SSE1 and MPD1; SSE1 and MPD2; SSE1 and EPS1; SSE1 and PDI1; SSE1
and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and UBC7; SSE1 and
DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2; SSE2 and ECM10;
SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2 and ERV2; SSE2
and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2 and
PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and UBC7;
SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10; SSB1
and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1 and
EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and PDI1;
SSB1 and DER1; SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7; SSB1
and DOA4; SSB1 and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2 and
MDJ2; SSB2 and ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and MPD1;
SSB2 and MPD2; SSB2 and EPS1; SSB2 and PDI1; SSB2 and DER1; SSB2
and DER3; SSB2 and HRD3; SSB2 and UBC7; SSB2 and DOA4; SSB2 and
HAC1; ECM10 and MDJ1; ECM10 and MDJ2; ECM10 and ERO1; ECM10 and
ERV2; ECM10 and EUG1; ECM10 and MPD1; ECM10 and MPD2; ECM10 and
EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and DER3; ECM10 and
HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and HAC1; MDJ1 and
MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1 and MPD1;
MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2
and DER3; MDJ2 and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and
HAC1; ERO1 and ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2;
ERO1 and EPS1; ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1
and HRD3; ERO1 and UBC7; ERO1 and DOA4; ERO1 and HAC1; ERV2 and
EUG1; ERV2 and MPD1; ERV2 and MPD2; ERV2 and EPS1; ERV2 and PDI1;
ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3; ERV2 and UBC7; ERV2
and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and
EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and
HRD3; MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1;
MPD2 and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2
and UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS I and PDI1; EPS1 and
DER1; EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0257] SSE1 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE2; JEM1 and SSB1; JEM1 and SSB2; JEM1
and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and ERO1; JEM1 and
ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1; LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE2; LHS1 and SSB1; LHS1
and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and MDJ2; LHS1 and
ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1; LHS1 and MPD2;
LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE2; SCJ1 and SSB1; SCJ1
and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and MDJ2; SCJ1 and
ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE2; KAR2 and SSB1; KAR2 and SSB2; KAR2
and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and ERO1; KAR2 and
ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2; KAR2 and EPS1;
KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE2;
SIL1 and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1 and MDJ1; SIL1
and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and EUG1; SIL1 and
MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1; SIL1 and DER1;
SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1
and HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and
SSA4; FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2; FKB2 and ECM10;
FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2 and ERV2; FKB2
and EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and EPS1; FKB2 and
PDI1; FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7;
FKB2 and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1
and SSA4; SSA1 and SSE2; SSA1 and SSB1; SSA1 and SSB2; SSA1 and
ECM10; SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1; SSA1 and ERV2;
SSA1 and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1
and PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and
UBC7; SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4;
SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2 and ECM10; SSA2
and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and ERV2; SSA2 and
EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2 and PDI1;
SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and UBC7; SSA2
and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE2; SSA3 and
SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3 and MDJ1; SSA3 and MDJ2;
SSA3 and ERO1; SSA3 and ERV2; SSA3 and EUG1; SSA3 and MPD1; SSA3
and MPD2; SSA3 and EPS1; SSA3 and PDI1; SSA3 and DER1; SSA3 and
DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3 and DOA4; SSA3 and HAC1;
SSA4 and SSE2; SSA4 and SSB1; SSA4 and SSB2; SSA4 and ECM10; SSA4
and MDJ1; SSA4 and MDJ2; SSA4 and ERO1; SSA4 and ERV2; SSA4 and
EUG1; SSA4 and MPD1; SSA4 and MPD2; SSA4 and EPS1; SSA4 and PDI1;
SSA4 and DER1; SSA4 and DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4
and DOA4; SSA4 and HAC1; SSE2 and SSB1; SSE2 and SSB2; SSE2 and
ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2 and ERV2;
SSE2 and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2
and PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and
UBC7; SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10;
SSB1 and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1
and EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and
PDI1; SSB1 and DER1; SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7;
SSB1 and DOA4; SSB1 and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2
and MDJ2; SSB2 and ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and
MPD1; SSB2 and MPD2; SSB2 and EPS1; SSB2 and PDI1; SSB2 and DER1;
SSB2 and DER3; SSB2 and HRD3; SSB2 and UBC7; SSB2 and DOA4; SSB2
and HAC1; ECM10 and MDJ1; ECM10 and MDJ2; ECM10 and ERO1; ECM10 and
ERV2; ECM10 and EUG1; ECM10 and MPD1; ECM10 and MPD2; ECM10 and
EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and DER3; ECM10 and
HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and HAC1; MDJ1 and
MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1 and MPD1;
MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2
and DER3; MDJ2 and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and
HAC1; ERO1 and ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2;
ERO1 and EPS1; ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1
and HRD3; ERO1 and UBC7; ERO1 and DOA4; ERO1 and HAC1; ERV2 and
EUG1; ERV2 and MPD1; ERV2 and MPD2; ERV2 and EPS1; ERV2 and PDI1;
ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3; ERV2 and UBC7; ERV2
and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and
EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and
HRD3; MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1;
MPD2 and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2
and UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and
DER1; EPS-1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0258] SSE2 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE1; JEM1 and SSB1; JEM1 and SSB2; JEM1
and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and ERO1; JEM1 and
ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1 LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSB1; LHS1
and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and MDJ2; LHS1 and
ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1; LHS1 and MPD2;
LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSB1; SCJ1
and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and MDJ2; SCJ1 and
ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSB1; KAR2 and SSB2; KAR2
and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and ERO1; KAR2 and
ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2; KAR2 and EPS1;
KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1;
SIL1 and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1 and MDJ1; SIL1
and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and EUG1; SIL1 and
MPD1; Sail and MPD2; SIL1 and EPS1; SIL1 and PDI1; SIL1 and DER1;
SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1
and HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and
SSA4; FKB2 and SSE1; FKB2 and SSB1; FKB2 and SSB2; FKB2 and ECM10;
FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2 and ERV2; FKB2
and EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and EPS1; FKB2 and
PDI1; FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7;
FKB2 and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1
and SSA4; SSA1 and SSE1; SSA1 and SSB1; SSA1 and SSB2; SSA1 and
ECM10; SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1; SSA1 and ERV2;
SSA1 and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1
and PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and
UBC7; SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4;
SSA2 and SSE1; SSA2 and SSB1; SSA2 and SSB2; SSA2 and ECM10; SSA2
and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and ERV2; SSA2 and
EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2 and PDI1;
SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and UBC7; SSA2
and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3 and
SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3 and MDJ1; SSA3 and MDJ2;
SSA3 and ERO1; SSA3 and ERV2; SSA3 and EUG1; SSA3 and MPD1; SSA3
and MPD2; SSA3 and EPS1; SSA3 and PDI1; SSA3 and DER1; SSA3 and
DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3 and DOA4; SSA3 and HAC1;
SSA4 and SSE1; SSA4 and SSB1; SSA4 and SSB2; SSA4 and ECM10; SSA4
and MDJ1; SSA4 and MDJ2; SSA4 and ERO1; SSA4 and ERV2; SSA4 and
EUG1; SSA4 and MPD1; SSA4 and MPD2; SSA4 and EPS1; SSA4 and PDI1;
SSA4 and DER1; SSA4 and DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4
and DOA4; SSA4 and HAC1; SSE1 and SSB1; SSE1 and SSB2; SSE1 and
ECM10; SSE1 and MDJ1; SSE1 and MDJ2; SSE1 and ERO1; SSE1 and ERV2;
SSE1 and EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1 and EPS1; SSE1
and PDI1; SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and
UBC7; SSE1 and DOA4; SSE1 and HAC1; SSB1 and SSB2; SSB1 and ECM10;
SSB1 and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1
and EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and
PDI1; SSB1 and DER1; SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7;
SSB1 and DOA4; SSB1 and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2
and MDJ2; SSB2 and ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and
MPD1; SSB2 and MPD2; SSB2 and EPS1; SSB2 and PDI1; SSB2 and DER1;
SSB2 and DER3; SSB2 and HRD3; SSB2 and UBC7; SSB2 and DOA4; SSB2
and HAC1; ECM10 and MDJ1; ECM10 and MDJ2; ECM10 and ERO1; ECM10 and
ERV2; ECM10 and EUG1; ECM10 and MPD1; ECM10 and MPD2; ECM10 and
EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and DER3; ECM10 and
HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and HAC1; MDJ1 and
MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1 and MPD1;
MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2
and DER3; MDJ2 and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and
HAC1; ERO1 and ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2;
ERO1 and EPS1; ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1
and HRD3; ERO1 and UBC7; ERO1 and DOA4; ERO1 and HAC1; ERV2 and
EUG1; ERV2 and MPD1; ERV2 and MPD2; ERV2 and EPS1; ERV2 and PDI1;
ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3; ERV2 and UBC7; ERV2
and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and
EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and
HRD3; MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1;
MPD2 and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2
and UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and
DER1; EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0259] SSB1 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB2; JEM1
and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and ERO1; JEM1 and
ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1; LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1
and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and MDJ2; LHS1 and
ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1; LHS1 and MPD2;
LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1
and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and MDJ2; SCJ1 and
ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB2; KAR2
and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and ERO1; KAR2 and
ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2; KAR2 and EPS1;
KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1;
SIL1 and SSE2; SIL1 and SSB2; SIL1 and ECM10; SIL1 and MDJ1; SIL1
and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and EUG1; SIL1 and
MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1; SIL1 and DER1;
SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1
and HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and
SSA4; FKB2 and SSE1; FKB2 and SSE2; FKB2 and SSB2; FKB2 and ECM10;
FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2 and ERV2; FKB2
and EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and EPS1; FKB2 and
PDI1; FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7;
FKB2 and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1
and SSA4; SSA1 and SSE1; SSA1 and SSE2; SSA1 and SSB2; SSA1 and
ECM10; SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1; SSA1 and ERV2;
SSA1 and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1
and PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and
UBC7; SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4;
SSA2 and SSE1; SSA2 and SSE2; SSA2 and SSB2; SSA2 and ECM10; SSA2
and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and ERV2; SSA2 and
EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2 and PDI1;
SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and UBC7; SSA2
and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3 and
SSE2; SSA3 and SSB2; SSA3 and ECM10; SSA3 and MDJ1; SSA3 and MDJ2;
SSA3 and ERO1; SSA3 and ERV2; SSA3 and EUG1; SSA3 and MPD1; SSA3
and MPD2; SSA3 and EPS1; SSA3 and PDI1; SSA3 and DER1; SSA3 and
DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3 and DOA4; SSA3 and HAC1;
SSA4 and SSE1; SSA4 and SSE2; SSA4 and SSB2; SSA4 and ECM10; SSA4
and MDJ1; SSA4 and MDJ2; SSA4 and ERO1; SSA4 and ERV2; SSA4 and
EUG1; SSA4 and MPD1; SSA4 and MPD2; SSA4 and EPS1; SSA4 and PDI1;
SSA4 and DER1; SSA4 and DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4
and DOA4; SSA4 and HAC1; SSE1 and SSE2; SSE1 and SSB2; SSE1 and
ECM10; SSE1 and MDJ1; SSE1 and MDJ2; SSE1 and ERO1; SSE1 and ERV2;
SSE1 and EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1 and EPS1; SSE1
and PDI1; SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and
UBC7; SSE1 and DOA4; SSE1 and HAC1; SSE2 and SSB2; SSE2 and ECM10;
SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2 and ERV2; SSE2
and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2 and
PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and UBC7;
SSE2 and DOA4; SSE2 and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2
and MDJ2; SSB2 and ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and
MPD1; SSB2 and MPD2; SSB2 and EPS1; SSB2 and PDI1; SSB2 and DER1;
SSB2 and DER3; SSB2 and HRD3; SSB2 and UBC7; SSB2 and DOA4; SSB2
and HAC1; ECM10 and MDJ1; ECM10 and MDJ2; ECM10 and ERO1; ECM10 and
ERV2; ECM10 and EUG1; ECM10 and MPD1; ECM10 and MPD2; ECM10 and
EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and DER3; ECM10 and
HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and HAC1; MDJ1 and
MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1 and MPD1;
MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2
and DER3; MDJ2 and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and
HAC1; ERO1 and ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2;
ERO1 and EPS1; ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1
and HRD3; ERO1 and UBC7; ERO1 and DOA4; ERO1 and HAC1; ERV2 and
EUG1; ERV2 and MPD1; ERV2 and MPD2; ERV2 and EPS1; ERV2 and PDI1;
ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3; ERV2 and UBC7; ERV2
and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and
EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and
HRD3; MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1;
MPD2 and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2
and UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and
DER1; EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0260] SSB2 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1
and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and ERO1; JEM1 and
ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1; LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1
and SSB1; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and MDJ2; LHS1 and
ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1; LHS1 and MPD2;
LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1
and SSB1; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and MDJ2; SCJ1 and
ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and ERO1; KAR2 and
ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2; KAR2 and EPS1;
KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1;
SIL1 and SSE2; SIL1 and SSB1; SIL1 and ECM10; SIL1 and MDJ1; SIL1
and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and EUG1; SILT and
MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1; SIL1 and DER1;
SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1
and HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and
SSA4; FKB2 and SSE1; FKB2 and SSE2; FKB2 and SSB1; FKB2 and ECM10;
FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2 and ERV2; FKB2
and EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and EPS1; FKB2 and
PDI1; FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7;
FKB2 and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1
and SSA4; SSA1 and SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and
ECM10; SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1; SSA1 and ERV2;
SSA1 and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1
and PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and
UBC7; SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4;
SSA2 and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and ECM10; SSA2
and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and ERV2; SSA2 and
EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2 and PDI1;
SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and UBC7; SSA2
and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3 and
SSE2; SSA3 and SSB1; SSA3 and ECM10; SSA3 and MDJ1; SSA3 and MDJ2;
SSA3 and ERO1; SSA3 and ERV2; SSA3 and EUG1; SSA3 and MPD1; SSA3
and MPD2; SSA3 and EPS1; SSA3 and PDI1; SSA3 and DER1; SSA3 and
DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3 and DOA4; SSA3 and HAC1;
SSA4 and SSE1; SSA4 and SSE2; SSA4 and SSB1; SSA4 and ECM10; SSA4
and MDJ1; SSA4 and MDJ2; SSA4 and ERO1; SSA4 and ERV2; SSA4 and
EUG1; SSA4 and MPD1; SSA4 and MPD2; SSA4 and EPS1; SSA4 and PDI1;
SSA4 and DER1; SSA4 and DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4
and DOA4; SSA4 and HAC1; SSE1 and SSE2; SSE1 and SSB1; SSE1 and
ECM10; SSE1 and MDJ1; SSE1 and MDJ2; SSE1 and ERO1; SSE1 and ERV2;
SSE1 and EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1 and EPS1; SSE1
and PDI1; SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and
UBC7; SSE1 and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and ECM10;
SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2 and ERV2; SSE2
and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2 and
PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and UBC7;
SSE2 and DOA4; SSE2 and HAC1; SSB1 and ECM10; SSB1 and MDJ1; SSB1
and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1 and EUG1; SSB1 and
MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and PDI1; SSB1 and DER1;
SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7; SSB1 and DOA4; SSB1
and HAC1; ECM10 and MDJ1; ECM10 and MDJ2; ECM10 and ERO1; ECM10 and
ERV2; ECM10 and EUG1; ECM10 and MPD1; ECM10 and MPD2; ECM10 and
EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and DER3; ECM10 and
HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and HAC1; MDJ1 and
MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1 and MPD1;
MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2
and DER3; MDJ2 and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and
HAC1; ERO1 and ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2;
ERO1 and EPS1; ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1
and HRD3; ERO1 and UBC7; ERO1 and DOA4; ERO1 and HAC1; ERV2 and
EUG1; ERV2 and MPD1; ERV2 and MPD2; ERV2 and EPS1; ERV2 and PDI1;
ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3; ERV2 and UBC7; ERV2
and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and
EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and
HRD3; MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1;
MPD2 and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2
and UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and
DER1; EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0261] ECM10 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; EMI and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1
and SSB2; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and ERO1; JEM1 and
ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEW and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1 LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1
and SSB1; LHS1 and SSB2; LHS1 and MDJ1; LHS1 and MDJ2; LHS1 and
ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1; LHS1 and MPD2;
LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SID; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1
and SSB1; SCJ1 and SSB2; SCJ1 and MDJ1; SCJ1 and MDJ2; SCJ1 and
ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSB2; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and ERO1; KAR2 and
ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2; KAR2 and EPS1;
KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1;
SIL1 and SSE2; SILT and SSB1; SIL1 and SSB2; SIL1 and MDJ1; SIL1
and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and EUG1; SIL1 and
MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1; SIL1 and DER1;
SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1
and HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and
SSA4; FKB2 and SSE1; FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2;
FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2 and ERV2; FKB2
and EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and EPS1; FKB2 and
PDI1; FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7;
FKB2 and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1
and SSA4; SSA1 and SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and
SSB2; SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1; SSA1 and ERV2;
SSA1 and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1
and PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and
UBC7; SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4;
SSA2 and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2
and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and ERV2; SSA2 and
EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2 and PDI1;
SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and UBC7; SSA2
and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3 and
SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and MDJ1; SSA3 and MDJ2;
SSA3 and ERO1; SSA3 and ERV2; SSA3 and EUG1; SSA3 and MPD1; SSA3
and MPD2; SSA3 and EPS1; SSA3 and PDI1; SSA3 and DER1; SSA3 and
DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3 and DOA4; SSA3 and HAC1;
SSA4 and SSE1; SSA4 and SSE2; SSA4 and SSB1; SSA4 and SSB2; SSA4
and MDJ1; SSA4 and MDJ2; SSA4 and ERO1; SSA4 and ERV2; SSA4 and
EUG1; SSA4 and MPD1; SSA4 and MPD2; SSA4 and EPS1; SSA4 and PDI1;
SSA4 and DER1; SSA4 and DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4
and DOA4; SSA4 and HAC1; SSE1 and SSE2; SSE1 and SSB1; SSE1 and
SSB2; SSE1 and MDJ1; SSE1 and MDJ2; SSE1 and ERO1; SSE1 and ERV2;
SSE1 and EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1 and EPS1; SSE1
and PDI1; SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and
UBC7; SSE1 and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2;
SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2 and ERV2; SSE2
and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2 and
PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and UBC7;
SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and MDJ1; SSB1
and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1 and EUG1; SSB1 and
MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and PDI1; SSB1 and DER1;
SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7; SSB1 and DOA4; SSB1
and HAC1; SSB2 and MDJ1; SSB2 and MDJ2; SSB2 and ERO1; SSB2 and
ERV2; SSB2 and EUG1; SSB2 and MPD1; SSB2 and MPD2; SSB2 and EPS1;
SSB2 and PDI1; SSB2 and DER1; SSB2 and DER3; SSB2 and HRD3; SSB2
and UBC7; SSB2 and DOA4; SSB2 and HAC1; MDJ1 and MDJ2; MDJ1 and
ERO1; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1 and MPD1; MDJ1 and MPD2;
MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1 and DER3; MDJ1
and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and HAC1; MDJ2 and
ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1; MDJ2 and MPD2;
MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2 and DER3; MDJ2
and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and HAC1; ERO1 and
ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2; ERO1 and EPS1;
ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1 and HRD3; ERO1
and UBC7; ERO1 and DOA4; ERO1 and HAC1; ERV2 and EUG1; ERV2 and
MPD1; ERV2 and MPD2; ERV2 and EPS1; ERV2 and PDI1; ERV2 and DER1;
ERV2 and DER3; ERV2 and HRD3; ERV2 and UBC7; ERV2 and DOA4; ERV2
and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and EPS1; EUG1 and
PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3; EUG1 and UBC7;
EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1 and EPS1; MPD1
and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and HRD3; MPD1 and
UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1; MPD2 and PDI1;
MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2 and UBC7; MPD2
and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and DER1; EPS1 and
DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4; EPS1 and HAC1;
PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1 and UBC7; PDI1
and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and HRD3; DER1 and
UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3; DER3 and UBC7;
DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3 and DOA4; HRD3
and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and HAC1.
[0262] MDJ1 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1
and SSB2; JEM1 and ECM10; JEM1 and MDJ2; JEM1 and ERO1; JEM1 and
ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1; LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1
and SSB1; LHS1 and SSB2; LHS1 and ECM10; LHS1 and MDJ2; LHS1 and
ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1; LHS1 and MPD2;
LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1
and SSB1; SCJ1 and SSB2; SCJ1 and ECM10; SCJ1 and MDJ2; SCJ1 and
ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSE2; KAR2 and ECM10; KAR2 and MDJ2; KAR2 and ERO1; KAR2 and
ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2; KAR2 and EPS1;
KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1;
SIL1 and SSE2; SIL1 and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1
and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and EUG1; SIL1 and
MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1; SIL1 and DER1;
SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1
and HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and
SSA4; FKB2 and SSE1; FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2;
FKB2 and ECM10; FKB2 and MDJ2; FKB2 and ERO1; FKB2 and ERV2; FKB2
and EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and EPS1; FKB2 and
PDI1; FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7;
FKB2 and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1
and SSA4; SSA1 and SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and
SSB2; SSA1 and ECM10; SSA1 and MDJ2; SSA1 and ERO1; SSA1 and ERV2;
SSA1 and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1
and PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and
UBC7; SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4;
SSA2 and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2
and ECM10; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and ERV2; SSA2 and
EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2 and PDI1;
SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and UBC7; SSA2
and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3 and
SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3 and MDJ2;
SSA3 and ERO1; SSA3 and ERV2; SSA3 and EUG1; SSA3 and MPD1; SSA3
and MPD2; SSA3 and EPS1; SSA3 and PDI1; SSA3 and DER1; SSA3 and
DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3 and DOA4; SSA3 and HAC1;
SSA4 and SSE1; SSA4 and SSE2; SSA4 and SSB1; SSA4 and SSB2; SSA4
and ECM10; SSA4 and MDJ2; SSA4 and ERO1; SSA4 and ERV2; SSA4 and
EUG1; SSA4 and MPD1; SSA4 and MPD2; SSA4 and EPS1; SSA4 and PDI1;
SSA4 and DER1; SSA4 and DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4
and DOA4; SSA4 and HAC1; SSE1 and SSE2; SSE1 and SSB1; SSE1 and
SSB2; SSE1 and ECM10; SSE1 and MDJ2; SSE1 and ERO1; SSE1 and ERV2;
SSE1 and EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1 and EPS1; SSE1
and PDI1; SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and
UBC7; SSE1 and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2;
SSE2 and ECM10; SSE2 and MDJ2; SSE2 and ERO1; SSE2 and ERV2; SSE2
and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2 and
PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and UBC7;
SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10; SSB1
and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1 and EUG1; SSB1 and
MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and PDI1; SSB1 and DER1;
SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7; SSB1 and DOA4; SSB1
and HAC1; SSB2 and ECM10; SSB2 and MDJ2; SSB2 and ERO1; SSB2 and
ERV2; SSB2 and EUG1; SSB2 and MPD1; SSB2 and MPD2; SSB2 and EPS1;
SSB2 and PDI1; SSB2 and DER1; SSB2 and DER3; SSB2 and HRD3; SSB2
and UBC7; SSB2 and DOA4; SSB2 and HAC1; ECM10 and MDJ2; ECM10 and
ERO1; ECM10 and ERV2; ECM10 and EUG1; ECM10 and MPD1; ECM10 and
MPD2; ECM10 and EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and
DER3; ECM10 and HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2
and DER3; MDJ2 and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and
HAC1; ERO1 and ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2;
ERO1 and EPS1; ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1
and HRD3; ERO1 and UBC7; ERO1 and DOA4; ERO1 and HAC1; ERV2 and
EUG1; ERV2 and MPD1; ERV2 and MPD2; ERV2 and EPS1; ERV2 and PDI1;
ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3; ERV2 and UBC7; ERV2
and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and
EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and
HRD3; MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1;
MPD2 and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2
and UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and
DER1; EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0263] MDJ2 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1
and SSB2; JEM1 and ECM10; JEM1 and MDJ1; JEM1 and ERO1; JEM1 and
ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1; LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1
and SSB1; LHS1 and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and
ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1; LHS1 and MPD2;
LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1
and SSB1; SCJ1 and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and
ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSB2; KAR2 and ECM10; KAR2 and MDJ1; KAR2 and ERO1; KAR2 and
ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2; KAR2 and EPS1;
KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1;
SIL1 and SSE2; SIL1 and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1
and MDJ1; SIL1 and ERO1; SIL1 and ERV2; SIL1 and EUG1; SIL1 and
MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1; SIL1 and DER1;
SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1
and HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and
SSA4; FKB2 and SSE1; FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2;
FKB2 and ECM10; FKB2 and MDJ1; FKB2 and ERO1; FKB2 and ERV2; FKB2
and EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and EPS1; FKB2 and
PDI1; FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7;
FKB2 and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1
and SSA4; SSA1 and SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and
SSB2; SSA1 and ECM10; SSA1 and MDJ1; SSA1 and ERO1; SSA1 and ERV2;
SSA1 and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1
and PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and
UBC7; SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4;
SSA2 and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2
and ECM10; SSA2 and MDJ1; SSA2 and ERO1; SSA2 and ERV2; SSA2 and
EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2 and PDI1;
SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and UBC7; SSA2
and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3 and
SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3 and MDJ1;
SSA3 and ERO1; SSA3 and ERV2; SSA3 and EUG1; SSA3 and MPD1; SSA3
and MPD2; SSA3 and EPS1; SSA3 and PDI1; SSA3 and DER1; SSA3 and
DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3 and DOA4; SSA3 and HAC1;
SSA4 and SSE1; SSA4 and SSE2; SSA4 and SSB1; SSA4 and SSB2; SSA4
and ECM10; SSA4 and MDJ1; SSA4 and ERO1; SSA4 and ERV2; SSA4 and
EUG1; SSA4 and MPD1; SSA4 and MPD2; SSA4 and EPS1; SSA4 and PDI1;
SSA4 and DER1; SSA4 and DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4
and DOA4; SSA4 and HAC1; SSE1 and SSE2; SSE1 and SSB1; SSE1 and
SSB2; SSE1 and ECM10; SSE1 and MDJ1; SSE1 and ERO1; SSE1 and ERV2;
SSE1 and EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1 and EPS1; SSE1
and PDI1; SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and
UBC7; SSE1 and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2;
SSE2 and ECM10; SSE2 and MDJ1; SSE2 and ERO1; SSE2 and ERV2; SSE2
and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2 and
PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and UBC7;
SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10; SSB1
and MDJ1; SSB1 and ERO1; SSB1 and ERV2; SSB1 and EUG1; SSB1 and
MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and PDI1; SSB1 and DER1;
SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7; SSB1 and DOA4; SSB1
and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2 and ERO1; SSB2 and
ERV2; SSB2 and EUG1; SSB2 and MPD1; SSB2 and MPD2; SSB2 and EPS1;
SSB2 and PDI1; SSB2 and DER1; SSB2 and DER3; SSB2 and HRD3; SSB2
and UBC7; SSB2 and DOA4; SSB2 and HAC1; ECM10 and MDJ1; ECM10 and
ERO1; ECM10 and ERV2; ECM10 and EUG1; ECM10 and MPD1; ECM10 and
MPD2; ECM10 and EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and
DER3; ECM10 and HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and
HAC1; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1 and MPD1;
MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; ERO1 and ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2;
ERO1 and EPS1; ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1
and HRD3; ERO1 and UBC7; ERO1 and DOA4; ERO1 and HAC1; ERV2 and
EUG1; ERV2 and MPD1; ERV2 and MPD2; ERV2 and EPS1; ERV2 and PDI1;
ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3; ERV2 and UBC7; ERV2
and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and
EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and
HRD3'; MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1;
MPD2 and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2
and UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and
DER1; EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0264] ERO1 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1
and SSB2; JEM1 and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and
ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1; LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1
and SSB1; LHS1 and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and
MDJ2; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1; LHS1 and MPD2;
LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1
and SSB1; SCJ1 and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and
MDJ2; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSB2; KAR2 and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and
ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2; KAR2 and EPS1;
KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and, SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1;
SIL1 and SSE2; SIL1 and SSB1; Slid and SSB2; SIL1 and ECM10; SIL1
and MDJ1; SIL1 and MDJ2; SIL1 and ERV2; SIL1 and EUG1; SIL1 and
MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1; SIL1 and DER1;
SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1
and HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and
SSA4; FKB2 and SSE1; FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2;
FKB2 and ECM10; FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERV2; FKB2
and EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and EPS1; FKB2 and
PDI1; FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7;
FKB2 and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1
and SSA4; SSA1 and SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and
SSB2; SSA1 and ECM10; SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERV2;
SSA1 and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1
and PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and
UBC7; SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4;
SSA2 and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2
and ECM10; SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERV2; SSA2 and
EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2 and PDI1;
SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and UBC7; SSA2
and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3 and
SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3 and MDJ1;
SSA3 MDJ2; SSA3 and ERV2; SSA3 and EUG1; SSA3 and MPD1; SSA3 and
MPD2; SSA3 and EPS1; SSA3 and PDI1; SSA3 and DER1; SSA3 and DER3;
SSA3 and HRD3; SSA3 and UBC7; SSA3 and DOA4; SSA3 and HAC1; SSA4
and SSE1; SSA4 and SSE2; SSA4 and SSB1; SSA4 and SSB2; SSA4 and
ECM10; SSA4 and MDJ1; SSA4 and MDJ2; SSA4 and ERV2; SSA4 and EUG1;
SSA4 and MPD1; SSA4 and MPD2; SSA4 and EPS1; SSA4 and PDI1; SSA4
and DER1; SSA4 and DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4 and
DOA4; SSA4 and HAC1; SSE1 and SSE2; SSE1 and SSB1; SSE1 and SSB2;
SSE1 and ECM10; SSE1 and MDJ1; SSE1 and MDJ2; SSE1 and ERV2; SSE1
and EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1 and EPS1; SSE1 and
PDI1; SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and UBC7;
SSE1 and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2; SSE2
and ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERV2; SSE2 and
EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2 and PDI1;
SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and UBC7; SSE2
and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10; SSB1 and
MDJ1; SSB1 and MDJ2; SSB1 and ERV2; SSB1 and EUG1; SSB1 and MPD1;
SSB1 and MPD2; SSB1 and EPS1; SSB1 and PDI1; SSB1 and DER1; SSB1
and DER3; SSB1 and HRD3; SSB1 and UBC7; SSB1 and DOA4; SSB1 and
HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2 and MDJ2; SSB2 and ERV2;
SSB2 and EUG1; SSB2 and MPD1; SSB2 and MPD2; SSB2 and EPS1; SSB2
and PDI1; SSB2 and DER1; SSB2 and DER3; SSB2 and HRD3; SSB2 and
UBC7; SSB2 and DOA4; SSB2 and HAC1; ECM10 and MDJ1; ECM10 and MDJ2;
ECM10 and ERV2; ECM10 and EUG1; ECM10 and MPD1; ECM10 and MPD2;
ECM10 and EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and DER3;
ECM10 and HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and HAC1;
MDJ1 and MDJ2; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1 and MPD1; MDJ1
and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1 and
DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and HAC1;
MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1; MDJ2 and MPD2; MDJ2
and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2 and DER3; MDJ2 and
HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and HAC1; ERV2 and EUG1;
ERV2 and MPD1; ERV2 and MPD2; ERV2 and EPS1; ERV2 and PDI1; ERV2
and DER1; ERV2 and DER3; ERV2 and HRD3; ERV2 and UBC7; ERV2 and
DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and EPS1;
EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3; EUG1
and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1 and
EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and HRD3;
MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1; MPD2
and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2 and
UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and DER1;
EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4; EPS1
and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1 and
UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and HRD3;
DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3; DER3
and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3 and
DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0265] ERV2 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1
and SSB2; JEM1 and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and
ERO1; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1; LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1
and SSB1; LHS1 and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and
MDJ2; LHS1 and ERO1; LHS1 and EUG1; LHS1 and MPD1; LHS1 and MPD2;
LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1
and SSB1; SCJ1 and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and
MDJ2; SCJ1 and ERO1; SCJ1 and EUG1; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSB2; KAR2 and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and
ERO1; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2; KAR2 and EPS1;
KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1;
SIL1 and SSE2; SIL1 and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1
and MDJ1; SIL1 and MDJ2; SIL1 and ERO1; SIL1 and EUG1; SIL1 and
MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1; SIL1 and DER1;
SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1
and HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and
SSA4; FKB2 and SSE1; FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2;
FKB2 and ECM10; FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2
and EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and EPS1; FKB2 and
PDI1; FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7;
FKB2 and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1
and SSA4; SSA1 and SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and
SSB2; SSA1 and ECM10; SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1;
SSA1 and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1
and PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and
UBC7; SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4;
SSA2 and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2
and ECM10; SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and
EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2 and PDI1;
SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and UBC7; SSA2
and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3 and
SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3 and MDJ1;
SSA3 and MDJ2; SSA3 and ERO1; SSA3 and EUG1; SSA3 and MPD1; SSA3
and MPD2; SSA3 and EPS1; SSA3 and PDI1; SSA3 and DER1; SSA3 and
DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3 and DOA4; SSA3 and HAC1;
SSA4 and SSE1; SSA4 and SSE2; SSA4 and SSB1; SSA4 and SSB2; SSA4
and ECM10; SSA4 and MDJ1; SSA4 and MDJ2; SSA4 and ERO1; SSA4 and
EUG1; SSA4 and MPD1; SSA4 and MPD2; SSA4 and EPS1; SSA4 and PDI1;
SSA4 and DER1; SSA4 and DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4
and DOA4; SSA4 and HAC1; SSE1 and SSE2; SSE1 and SSB1; SSE1 and
SSB2; SSE1 and ECM10; SSE1 and MDJ1; SSE1 and MDJ2; SSE1 and ERO1;
SSE1 and EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1 and EPS1; SSE1
and PDI1; SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and
UBC7; SSE1 and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2;
SSE2 and ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2
and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2 and
PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and UBC7;
SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10; SSB1
and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and EUG1; SSB1 and
MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and PDI1; SSB1 and DER1;
SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7; SSB1 and DOA4; SSB1
and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2 and MDJ2; SSB2 and
ERO1; SSB2 and EUG1; SSB2 and MPD1; SSB2 and MPD2; SSB2 and EPS1;
SSB2 and PDI1; SSB2 and DER1; SSB2 and DER3; SSB2 and HRD3; SSB2
and UBC7; SSB2 and DOA4; SSB2 and HAC1; ECM10 and MDJ1; ECM10 and
MDJ2; ECM10 and ERO1; ECM10 and EUG1; ECM10 and MPD1; ECM10 and
MPD2; ECM10 and EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and
DER3; ECM10 and HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and
MeI; MDJ1 and MDJ2; MDJ1 and ERO1; MDJ1 and EUG1; MDJ1 and MPD1;
MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and EUG1; MDJ2 and MPD1; MDJ2 and MPD2;
MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2 and DER3; MDJ2
and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and HAC1; ERO1 and
EUG1; ERO1 and MPD1; ERO1 and MPD2; ERO1 and EPS1; ERO1 and PDI1;
ERO1 and DER1; ERO1 and DER3; ERO1 and HRD3; ERO1 and UBC7; ERO1
and DOA4; ERO1 and HAC1; EUG1 and MPD1; EUG1 and MPD2; EUG1 and
EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and
HRD3; MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1;
MPD2 and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2
and UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and
DER1; EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0266] EUG1 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1
and SSB2; JEM1 and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and
ERO1; JEM1 and ERV2; JEM1 and MPD1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1; LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1
and SSB1; LHS1 and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and
MDJ2; LHS1 and ERO1; LHS1 and ERV2; LHS1 and MPD1; LHS1 and MPD2;
LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1
and SSB1; SCJ1 and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and
MDJ2; SCJ1 and ERO1; SCJ1 and ERV2; SCJ1 and MPD1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSB2; KAR2 and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and
ERO1; KAR2 and ERV2; KAR2 and MPD1; KAR2 and MPD2; KAR2 and EPS1;
KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1;
SIL1 and SSE2; SIL1 and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1
and MDJ1; SIL1 and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and
MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1; SIL1 and DER1;
SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1
and HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and
SSA4; FKB2 and SSE1; FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2;
FKB2 and ECM10; FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2
and ERV2; FKB2 and MPD1; FKB2 and MPD2; FKB2 and EPS1; FKB2 and
PDI1; FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7;
FKB2 and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1
and SSA4; SSA1 and SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and
SSB2; SSA1 and ECM10; SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1;
SSA1 and ERV2; SSA1 and MPD1; SSA1 and MPD2; SSA1 and EPS1; SSA1
and PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and
UBC7; SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4;
SSA2 and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2
and ECM10; SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and
ERV2; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2 and PDI1;
SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and UBC7; SSA2
and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3 and
SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3 and MDJ1;
SSA3 and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and MPD1; SSA3
and MPD2; SSA3 and EPS1; SSA3 and PDI1; SSA3 and DER1; SSA3 and
DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3 and DOA4; SSA3 and HAC1;
SSA4 and SSE1; SSA4 and SSE2; SSA4 and SSB1; SSA4 and SSB2; SSA4
and ECM10; SSA4 and MDJ1; SSA4 and MDJ2; SSA4 and ERO1; SSA4 and
ERV2; SSA4 and MPD1; SSA4 and MPD2; SSA4 and EPS1; SSA4 and PDI1;
SSA4 and DER1; SSA4 and DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4
and DOA4; SSA4 and HAC1; SSE1 and SSE2; SSE1 and SSB1; SSE1 and
SSB2; SSE1 and ECM10; SSE1 and MDJ1; SSE1 and MDJ2; SSE1 and ERO1;
SSE1 and ERV2; SSE1 and MPD1; SSE1 and MPD2; SSE1 and EPS1; SSE1
and PDI1; SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and
UBC7; SSE1 and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2;
SSE2 and ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2
and ERV2; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1; SSE2 and
PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and UBC7;
SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10; SSB1
and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1 and
MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and PDI1; SSB1 and DER1;
SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7; SSB1 and DOA4; SSB1
and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2 and MDJ2; SSB2 and
ERO1; SSB2 and ERV2; SSB2 and MPD1; SSB2 and MPD2; SSB2 and EPS1;
SSB2 and PDI1; SSB2 and DER1; SSB2 and DER3; SSB2 and HRD3; SSB2
and UBC7; SSB2 and DOA4; SSB2 and HAC1; ECM10 and MDJ1; ECM10 and
MDJ2; ECM10 and ERO1; ECM10 and ERV2; ECM10 and MPD1; ECM10 and
MPD2; ECM10 and EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and
DER3; ECM10 and HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and
HAC1; MDJ1 and MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and MPD1;
MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and MPD1; MDJ2 and MPD2;
MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2 and DER3; MDJ2
and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and HAC1; ERO1 and
ERV2; ERO1 and MPD1; ERO1 and MPD2; ERO1 and EPS1; ERO1 and PDI1;
ERO1 and DER1; ERO1 and DER3; ERO1 and HRD3; ERO1 and UBC7; ERO1
and DOA4; ERO1 and HAC1; ERV2 and MPD1; ERV2 and MPD2; ERV2 and
EPS1; ERV2 and PDI1; ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3;
ERV2 and UBC7; ERV2 and DOA4; ERV2 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and
HRD3; MPD1 and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1;
MPD2 and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2
and UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and
DER1; EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0267] MPD1 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1
and SSB2; JEM1 and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and
ERO1; JEW and ERV2; JEM1 and EUG1; JEM1 and MPD2; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1 LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1
and SSB1; LHS1 and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and
MDJ2; LHS1 and ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD2;
LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1
and SSB1; SCJ1 and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and
MDJ2; SCJ1 and ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD2;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSB2; KAR2 and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and
ERO1; KAR2 and ERV2; KAR2 and EUG1; KAR2 and MPD2; KAR2 and EPS1;
KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1;
SIL1 and SSE2; SIL1 and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1
and MDJ1; SIL1 and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and
EUG1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1; SIL1 and DER1;
SEM and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1 and
HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and SSA4;
FKB2 and SSE1; FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2; FKB2
and ECM10; FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2 and
ERV2; FKB2 and EUG1; FKB2 and MPD2; FKB2 and EPS1; FKB2 and PDI1;
FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7; FKB2
and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1 and
SSA4; SSA1 and SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and SSB2;
SSA1 and ECM10; SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1; SSA1
and ERV2; SSA1 and EUG1; SSA1 and MPD2; SSA1 and EPS1; SSA1 and
PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and UBC7;
SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4; SSA2
and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2 and
ECM10; SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and ERV2;
SSA2 and EUG1; SSA2 and MPD2; SSA2 and EPS1; SSA2 and PDI1; SSA2
and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and UBC7; SSA2 and
DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3 and SSE2;
SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3 and MDJ1; SSA3
and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and EUG1; SSA3 and
MPD2; SSA3 and EPS1; SSA3 and PDI1; SSA3 and DER1; SSA3 and DER3;
SSA3 and HRD3; SSA3 and UBC7; SSA3 and DOA4; SSA3 and HAC1; SSA4
and SSE1; SSA4 and SSE2; SSA4 and SSB1; SSA4 and SSB2; SSA4 and
ECM10; SSA4 and MDJ1; SSA4 and MDJ2; SSA4 and ERO1; SSA4 and ERV2;
SSA4 and EUG1; SSA4 and MPD2; SSA4 and EPS1; SSA4 and PDI1; SSA4
and DER1; SSA4 and DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4 and
DOA4; SSA4 and HAC1; SSE1 and SSE2; SSE1 and SSB1; SSE1 and SSB2;
SSE1 and ECM10; SSE1 and MDJ1; SSE1 and MDJ2; SSE1 and ERO1; SSE1
and ERV2; SSE1 and EUG1; SSE1 and MPD2; SSE1 and EPS1; SSE1 and
PDI1; SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and UBC7;
SSE1 and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2; SSE2
and ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2 and
ERV2; SSE2 and EUG1; SSE2 and MPD2; SSE2 and EPS1; SSE2 and PDI1;
SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and UBC7; SSE2
and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10; SSB1 and
MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1 and EUG1;
SSB1 and MPD2; SSB1 and EPS1; SSB1 and PDI1; SSB1 and DER1; SSB1
and DER3; SSB1 and HRD3; SSB1 and UBC7; SSB1 and DOA4; SSB1 and
HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2 and MDJ2; SSB2 and ERO1;
SSB2 and ERV2; SSB2 and EUG1; SSB2 and MPD2; SSB2 and EPS1; SSB2
and PDI1; SSB2 and DER1; SSB2 and DER3; SSB2 and HRD3; SSB2 and
UBC7; SSB2 and DOA4; SSB2 and HAC1; ECM10 and MDJ1; ECM10 and MDJ2;
ECM10 and ERO1; ECM10 and ERV2; ECM10 and EUG1; ECM10 and MPD2;
ECM10 and EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and DER3;
ECM10 and HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and HAC1;
MDJ1 and MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1
and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1 and
DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and HAC1;
MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD2; MDJ2
and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2 and DER3; MDJ2 and
HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and HAC1; ERO1 and ERV2;
ERO1 and EUG1; ERO1 and MPD2; ERO1 and EPS1; ERO1 and PDI1; ERO1
and DER1; ERO1 and DER3; ERO1 and HRD3; ERO1 and UBC7; ERO1 and
DOA4; ERO1 and HAC1; ERV2 and EUG1; ERV2 and MPD2; ERV2 and EPS1;
ERV2 and PDI1; ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3; ERV2
and UBC7; ERV2 and DOA4; ERV2 and HAC1; EUG1 and MPD2; EUG1 and
EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD2 and EPS1; MPD2
and PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2 and
UBC7; MPD2 and DOA4; MPD2 and HAC1; EPS1 and PDI1; EPS1 and DER1;
EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4; EPS1
and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1 and
UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and HRD3;
DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3; DER3
and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3 and
DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0268] MPD2 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1
and SSB2; JEM1 and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and
ERO1; JEM1 and ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and EPS1;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and MIA; LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1
and SSB1; LHS1 and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and
MDJ2; LHS1 and ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1;
LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1
and SSB1; SCJ1 and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and
MDJ2; SCJ1 and ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1;
SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSB2; KAR2 and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and
ERO1; KAR2 and ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and EPS1;
KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1;
SIL1 and SSE2; SIL1 and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1
and MDJ1; SIL1 and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and
EUG1; SIL1 and MPD1; SIL1 and EPS1; SIL1 and PDI1; SIL1 and DER1;
SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1
and HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and
SSA4; FKB2 and SSE1; FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2;
FKB2 and ECM10; FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2
and ERV2; FKB2 and EUG1; FKB2 and MPD1; FKB2 and EPS1; FKB2 and
PDI1; FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7;
FKB2 and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1
and SSA4; SSA1 and SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and
SSB2; SSA1 and ECM10; SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1;
SSA1 and ERV2; SSA1 and EUG1; SSA1 and MPD1; SSA1 and EPS1; SSA1
and PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and
UBC7; SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4;
SSA2 and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2
and ECM10; SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and
ERV2; SSA2 and EUG1; SSA2 and MPD1; SSA2 and EPS1; SSA2 and PDI1;
SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and UBC7; SSA2
and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3 and
SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3 and MDJ1;
SSA3 and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and EUG1; SSA3
and MPD1; SSA3 and EPS1; SSA3 and PDI1; SSA3 and DER1; SSA3 and
DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3 and DOA4; SSA3 and HAC1;
SSA4 and SSE1; SSA4 and SSE2; SSA4 and SSB1; SSA4 and SSB2; SSA4
and ECM10; SSA4 and MDJ1; SSA4 and MDJ2; SSA4 and ERO1; SSA4 and
ERV2; SSA4 and EUG1; SSA4 and MPD1; SSA4 and EPS1; SSA4 and PDI1;
SSA4 and DER1; SSA4 and DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4
and DOA4; SSA4 and HAC1; SSE1 and SSE2; SSE1 and SSB1; SSE1 and
SSB2; SSE1 and ECM10; SSE1 and MDJ1; SSE1 and MDJ2; SSE1 and ERO1;
SSE1 and ERV2; SSE1 and EUG1; SSE1 and MPD1; SSE1 and EPS1; SSE1
and PDI1; SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and
UBC7; SSE1 and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2;
SSE2 and ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2
and ERV2; SSE2 and EUG1; SSE2 and MPD1; SSE2 and EPS1; SSE2 and
PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and UBC7;
SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10; SSB1
and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1 and
EUG1; SSB1 and MPD1; SSB1 and EPS1; SSB1 and PDI1; SSB1 and DER1;
SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7; SSB1 and DOA4; SSB1
and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2 and MDJ2; SSB2 and
ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and MPD1; SSB2 and EPS1;
SSB2 and PDI1; SSB2 and DER1; SSB2 and DER3; SSB2 and HRD3; SSB2
and UBC7; SSB2 and DOA4; SSB2 and HAC1; ECM10 and MDJ1; ECM10 and
MDJ2; ECM10 and ERO1; ECM10 and ERV2; ECM10 and EUG1; ECM10 and
MPD1; ECM10 and EPS1; ECM10 and PDI1; ECM10 and DER1; ECM10 and
DER3; ECM10 and HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and
HAC1; MDJ1 and MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1;
MDJ1 and MPD1; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2 and DER3; MDJ2
and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and HAC1; ERO1 and
ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and EPS1; ERO1 and PDI1;
ERO1 and DER1; ERO1 and DER3; ERO1 and HRD3; ERO1 and UBC7; ERO1
and DOA4; ERO1 and HAC1; ERV2 and EUG1; ERV2 and MPD1; ERV2 and
EPS1; ERV2 and PDI1; ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3;
ERV2 and UBC7; ERV2 and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1
and EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and
HRD3; EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and EPS1;
MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and HRD3; MPD1
and UBC7; MPD1 and DOA4; MPD1 and HAC1; EPS1 and PDI1; EPS1 and
DER1; EPS1 and DER3; EPS1 and HRD3; EPS1 and UBC7; EPS1 and DOA4;
EPS1 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0269] EPS1 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1
and SSB2; JEM1 and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and
ERO1; JEM1 and ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2;
JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1; LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1
and SSB1; LHS1 and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and
MDJ2; LHS1 and ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1;
LHS1 and MPD2; LHS1 and PDI1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1
and SSB1; SCJ1 and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and
MDJ2; SCJ1 and ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1;
SCJ1 and MPD2; SCJ1 and PDI1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSB2; KAR2 and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and
ERO1; KAR2 and ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2;
KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1;
SIL1 and SSE2; SIL1 and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1
and MDJ1; SIL1 and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and
EUG1; SIL1 and MPD1; SIL1 and MPD2; SIL1 and PDI1; SIL1 and DER1;
SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1
and HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and
SSA4; FKB2 and SSE1; FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2;
FKB2 and ECM10; FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2
and ERV2; FKB2 and EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and
PDI1; FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7;
FKB2 and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1
and SSA4; SSA1 and SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and
SSB2; SSA1 and ECM10; SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1;
SSA1 and ERV2; SSA1 and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1
and PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and
UBC7; SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4;
SSA2 and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2
and ECM10; SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and
ERV2; SSA2 and EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and PDI1;
SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and UBC7; SSA2
and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3 and
SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3 and MDJ1;
SSA3 and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and EUG1; SSA3
and MPD1; SSA3 and MPD2; SSA3 and PDI1; SSA3 and DER1; SSA3 and
DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3 and DOA4; SSA3 and HAC1;
SSA4 and SSE1; SSA4 and SSE2; SSA4 and SSB1; SSA4 and SSB2; SSA4
and ECM10; SSA4 and MDJ1; SSA4 and MDJ2; SSA4 and ERO1; SSA4 and
ERV2; SSA4 and EUG1; SSA4 and MPD1; SSA4 and MPD2; SSA4 and PDI1;
SSA4 and DER1; SSA4 and DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4
and DOA4; SSA4 and HAC1; SSE1 and SSE2; SSE1 and SSB1; SSE1 and
SSB2; SSE1 and ECM10; SSE1 and MDJ1; SSE1 and MDJ2; SSE1 and ERO1;
SSE1 and ERV2; SSE1 and EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1
and PDI1; SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and
UBC7; SSE1 and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2;
SSE2 and ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2
and ERV2; SSE2 and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and
PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and UBC7;
SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10; SSB1
and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1 and
EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and PDI1; SSB1 and DER1;
SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7; SSB1 and DOA4; SSB1
and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2 and MDJ2; SSB2 and
ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and MPD1; SSB2 and MPD2;
SSB2 and PDI1; SSB2 and DER1; SSB2 and DER3; SSB2 and HRD3; SSB2
and UBC7; SSB2 and DOA4; SSB2 and HAC1; ECM10 and MDJ1; ECM10 and
MDJ2; ECM10 and ERO1; ECM10 and ERV2; ECM10 and EUG1; ECM10 and
MPD1; ECM10 and MPD2; ECM10 and PDI1; ECM10 and DER1; ECM10 and
DER3; ECM10 and HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and
HAM; MDJ1 and MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1;
MDJ1 and MPD1; MDJ1 and MPD2; MDJ1 and PDI1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and PDI1; MDJ2 and DER1; MDJ2 and DER3; MDJ2
and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and HAC1; ERO1 and
ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2; ERO1 and PDI1;
ERO1 and DER1; ERO1 and DER3; ERO1 and HRD3; ERO1 and UBC7; ERO1
and DOA4; ERO1 and HAC1; ERV2 and EUG1; ERV2 and MPD1; ERV2 and
MPD2; ERV2 and PDI1; ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3;
ERV2 and UBC7; ERV2 and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1
and MPD2; EUG1 and PDI1; EUG1 and DER1; EUG1 and DER3; EUG1 and
HRD3; EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2;
MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1 and HRD3; MPD1
and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and PDI1; MPD2 and
DER1; MPD2 and DER3; MPD2 and HRD3; MPD2 and UBC7; MPD2 and DOA4;
MPD2 and HAC1; PDI1 and DER1; PDI1 and DER3; PDI1 and HRD3; PDI1
and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0270] PDI1 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1
and SSB2; JEM1 and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and
ERO1; JEM1 and ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2;
JEM1 and EPS1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1 LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1
and SSB1; LHS1 and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and
MDJ2; LHS1 and ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1;
LHS1 and MPD2; LHS1 and EPS1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1
and SSB1; SCJ1 and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and
MDJ2; SCJ1 and ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1;
SCJ1 and MPD2; SCJ1 and EPS1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSB2; KAR2 and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and
ERO1; KAR2 and ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2;
KAR2 and EPS1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1;
SIL1 and SSE2; SIL1 and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1
and MDJ1; SIL1 and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and
EUG1; SIL1 and MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and DER1;
SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1
and HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and
SSA4; FKB2 and SSE1; FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2;
FKB2 and ECM10; FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2
and ERV2; FKB2 and EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and
EPS1; FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7;
FKB2 and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1
and SSA4; SSA1 and SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and
SSB2; SSA1 and ECM10; SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1;
SSA1 and ERV2; SSA1 and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1
and EPS1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and
UBC7; SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4;
SSA2 and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2
and ECM10; SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and
ERV2; SSA2 and EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1;
SSA2, and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and UBC7; SSA2
and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3 and
SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3 and MDJ1;
SSA3 and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and EUG1; SSA3
and MPD1; SSA3 and MPD2; SSA3 and EPS1; SSA3 and DER1; SSA3 and
DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3 and DOA4; SSA3 and HAC1;
SSA4 and SSE1; SSA4 and SSE2; SSA4 and SSB1; SSA4 and SSB2; SSA4
and ECM10; SSA4 and MDJ1; SSA4 and MDJ2; SSA4 and ERO1; SSA4 and
ERV2; SSA4 and EUG1; SSA4 and MPD1; SSA4 and MPD2; SSA4 and EPS1;
SSA4 and DER1; SSA4 and DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4
and DOA4; SSA4 and HAC1; SSE1 and SSE2; SSE1 and SSB1; SSE1 and
SSB2; SSE1 and ECM10; SSE1 and MDJ1; SSE1 and MDJ2; SSE1 and ERO1;
SSE1 and ERV2; SSE1 and EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1
and EPS1; SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and
UBC7; SSE1 and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2;
SSE2 and ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2
and ERV2; SSE2 and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and
EPS1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and UBC7;
SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10; SSB1
and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1 and
EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and DER1;
SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7; SSB1 and DOA4; SSB1
and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2 and MDJ2; SSB2 and
ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and MPD1; SSB2 and MPD2;
SSB2 and EPS1; SSB2 and DER1; SSB2 and DER3; SSB2 and HRD3; SSB2
and UBC7; SSB2 and DOA4; SSB2 and HAC1; ECM10 and MDJ1; ECM10 and
MDJ2; ECM10 and ERO1; ECM10 and ERV2; ECM10 and EUG1; ECM10 and
MPD1; ECM10 and MPD2; ECM10 and EPS1; ECM10 and DER1; ECM10 and
DER3; ECM10 and HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and
HAC1; MDJ1 and MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1;
MDJ1 and MPD1; MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and DER1; MDJ2 and DER3; MDJ2
and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and HAC1; ERO1 and
ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2; ERO1 and EPS1;
ERO1 and DER1; ERO1 and DER3; ERO1 and HRD3; ERO1 and UBC7; ERO1
and DOA4; ERO1 and HAC1; ERV2 and EUG1; ERV2 and MPD1; ERV2 and
MPD2; ERV2 and EPS1; ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3;
ERV2 and UBC7; ERV2 and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1
and MPD2; EUG1 and EPS1; EUG1 and DER1; EUG1 and DER3; EUG1 and
HRD3; EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2;
MPD1 and EPS1; MPD1 and DER1; MPD1 and DER3; MPD1 and HRD3; MPD1
and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1; MPD2 and
DER1; MPD2 and DER3; MPD2 and HRD3; MPD2 and UBC7; MPD2 and DOA4;
MPD2 and HAC1; EPS1 and DER1; EPS1 and DER3; EPS1 and HRD3; EPS1
and UBC7; EPS1 and DOA4; EPS1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0271] DER1 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1
and SSB2; JEM1 and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and
ERO1; JEM1 and ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2;
JEM1 and EPS1; JEM1 and DER1; JEM1 and DER3; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1 LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1
and SSB1; LHS1 and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and
MDJ2; LHS1 and ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1;
LHS1 and MPD2; LHS1 and EPS1; LHS1 and DER1; LHS1 and DER3; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1
and SSB1; SCJ1 and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and
MDJ2; SCJ1 and ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1;
SCJ1 and MPD2; SCJ1 and EPS1; SCJ1 and DER1; SCJ1 and DER3; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSB2; KAR2 and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and
ERO1; KAR2 and ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2;
KAR2 and EPS1; KAR2 and DER1; KAR2 and DER3; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1;
SIL1 and SSE2; SIL1 and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1
and MDJ1; SIL1 and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and
EUG1; SIL1 and MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and DER1;
SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1
and HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and
SSA4; FKB2 and SSE1; FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2;
FKB2 and ECM10; FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2
and ERV2; FKB2 and EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and
EPS1; FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3; FKB2 and UBC7;
FKB2 and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1
and SSA4; SSA1 and SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and
SSB2; SSA1 and ECM10; SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1;
SSA1 and ERV2; SSA1 and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1
and EPS1; SSA1 and DER1; SSA1 and DER3; SSA1 and HRD3; SSA1 and
UBC7; SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4;
SSA2 and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2
and ECM10; SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and
ERV2; SSA2 and EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1;
SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2 and UBC7; SSA2
and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3 and
SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3 and MDJ1;
SSA3 and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and EUG1; SSA3
and MPD1; SSA3 and MPD2; SSA3 and EPS1; SSA3 and DER1; SSA3 and
DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3 and DOA4; SSA3 and HAC1;
SSA4 and SSE1; SSA4 and SSE2; SSA4 and SSB1; SSA4 and SSB2; SSA4
and ECM10; SSA4 and MDJ1; SSA4 and MDJ2; SSA4 and ERO1; SSA4 and
ERV2; SSA4 and EUG1; SSA4 and MPD1; SSA4 and MPD2; SSA4 and EPS1;
SSA4 and DER1; SSA4 and DER3; SSA4 and HRD3; SSA4 and UBC7; SSA4
and DOA4; SSA4 and HAC1; SSE1 and SSE2; SSE1 and SSB1; SSE1 and
SSB2; SSE1 and ECM10; SSE1 and MDJ1; SSE1 and MDJ2; SSE1 and ERO1;
SSE1 and ERV2; SSE1 and EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1
and EPS1; SSE1 and DER1; SSE1 and DER3; SSE1 and HRD3; SSE1 and
UBC7; SSE1 and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2;
SSE2 and ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2
and ERV2; SSE2 and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and
EPS1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3; SSE2 and UBC7;
SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10; SSB1
and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1 and
EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and DER1;
SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7; SSB1 and DOA4; SSB1
and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2 and MDJ2; SSB2 and
ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and MPD1; SSB2 and MPD2;
SSB2 and EPS1; SSB2 and DER1; SSB2 and DER3; SSB2 and HRD3; SSB2
and UBC7; SSB2 and DOA4; SSB2 and HAC1; ECM10 and MDJ1; ECM10 and
MDJ2; ECM10 and ERO1; ECM10 and ERV2; ECM10 and EUG1; ECM10 and
MPD1; ECM10 and MPD2; ECM10 and EPS1; ECM10 and DER1; ECM10 and
DER3; ECM10 and HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and
HAC1; MDJ1 and MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1;
MDJ1 and MPD1; MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and DER1; MDJ1
and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and DER1; MDJ2 and DER3; MDJ2
and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and HAC1; ERO1 and
ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2; ERO1 and EPS1;
ERO1 and DER1; ERO1 and DER3; ERO1 and HRD3; ERO1 and UBC7; ERO1
and DOA4; ERO1 and HAC1; ERV2 and EUG1; ERV2 and MPD1; ERV2 and
MPD2; ERV2 and EPS1; ERV2 and DER1; ERV2 and DER3; ERV2 and HRD3;
ERV2 and UBC7; ERV2 and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1
and MPD2; EUG1 and EPS1; EUG1 and DER1; EUG1 and DER3; EUG1 and
HRD3; EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2;
MPD1 and EPS1; MPD1 and DER1; MPD1 and DER3; MPD1 and HRD3; MPD1
and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1; MPD2 and
DER1; MPD2 and DER3; MPD2 and HRD3; MPD2 and UBC7; MPD2 and DOA4;
MPD2 and HAC1; EPS1 and DER1; EPS1 and DER3; EPS1 and HRD3; EPS1
and UBC7; EPS1 and DOA4; EPS1 and HAC1; DER1 and DER3; DER1 and
HRD3; DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3;
DER3 and UBC7; DER3 and DOA4; DER3 and HAC1; HRD3 and UBC7; HRD3
and DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0272] DER3 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1
and SSB2; JEM1 and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and
ERO1; JEM1 and ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2;
JEM1 and EPS1; JEM1 and PDI1; JEM1 and DER1; JEM1 and HRD3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1; LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1
and SSB1; LHS1 and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and
MDJ2; LHS1 and ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1;
LHS1 and MPD2; LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1
and HRD3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1
and SSB1; SCJ1 and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and
MDJ2; SCJ1 and ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1;
SCJ1 and MPD2; SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1
and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSB2; KAR2 and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and
ERO1; KAR2 and ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2;
KAR2 and EPS1; KAR2 and PDI1; KAR2 and DER1; KAR2 and HRD3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; and FKB2; SIL1 and SSA1;
SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1; SIL1
and SSE2; SIL1 and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1 and
MDJ1; SIL1 and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and EUG1;
SIL1 and MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1; SIL1
and DER1; SIL1 and HRD3; SIL1 and UBC7; SIL1 and DOA4; SIL1 and
HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and SSA4;
FKB2 and SSE1; FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2; FKB2
and ECM10; FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2 and
ERV2; FKB2 and EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and EPS1;
FKB2 and PDI1; FKB2 and DER1; FKB2 and HRD3; FKB2 and UBC7; FKB2
and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1 and
SSA4; SSA1 and SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and SSB2;
SSA1 and ECM10; SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1; SSA1
and ERV2; SSA1 and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1 and
EPS1; SSA1 and PDI1; SSA1 and DER1; SSA1 and HRD3; SSA1 and UBC7;
SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4; SSA2
and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2 and
ECM10; SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and ERV2;
SSA2 and EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1; SSA2
and PDI1; SSA2 and DER1; SSA2 and HRD3; SSA2 and UBC7; SSA2 and
DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3 and SSE2;
SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3 and MDJ1; SSA3
and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and EUG1; SSA3 and
MPD1; SSA3 and MPD2; SSA3 and EPS1; SSA3 and PDI1; SSA3 and DER1;
SSA3 and HRD3; SSA3 and UBC7; SSA3 and DOA4; SSA3 and HAC1; SSA4
and SSE1; SSA4 and SSE2; SSA4 and SSB1; SSA4 and SSB2; SSA4 and
ECM10; SSA4 and MDJ1; SSA4 and MDJ2; SSA4 and ERO1; SSA4 and ERV2;
SSA4 and EUG1; SSA4 and MPD1; SSA4 and MPD2; SSA4 and EPS1; SSA4
and PDI1; SSA4 and DER1; SSA4 and HRD3; SSA4 and UBC7; SSA4 and
DOA4; SSA4 and HAC1; SSE1 and SSE2; SSE1 and SSB1; SSE1 and SSB2;
SSE1 and ECM10; SSE1 and MDJ1; SSE1 and MDJ2; SSE1 and ERO1; SSE1
and ERV2; SSE1 and EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1 and
EPS1; SSE1 and PDI1; SSE1 and DER1; SSE1 and HRD3; SSE1 and UBC7;
SSE1 and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2; SSE2
and ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2 and
ERV2; SSE2 and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and EPS1;
SSE2 and PDI1; SSE2 and DER1; SSE2 and HRD3; SSE2 and UBC7; SSE2
and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10; SSB1 and
MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1 and EUG1;
SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and PDI1; SSB1
and DER1; SSB1 and HRD3; SSB1 and UBC7; SSB1 and DOA4; SSB1 and
HAC1, SSB2 and ECM10; SSB2 and MDJ1; SSB2 and MDJ2; SSB2 and ERO1;
SSB2 and ERV2; SSB2 and EUG1; SSB2 and MPD1; SSB2 and MPD2; SSB2
and EPS1; SSB2 and PDI1; SSB2 and DER1; SSB2 and HRD3; SSB2 and
UBC7; SSB2 and DOA4; SSB2 and HAC1; ECM10 and MDJ1; ECM10 and MDJ2;
ECM10 and ERO1; ECM10 and ERV2; ECM10 and EUG1; ECM10 and MPD1;
ECM10 and MPD2; ECM10 and EPS1; ECM10 and PDI1; ECM10 and DER1;
ECM10 and HRD3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and HAC1;
MDJ1 and MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1; MDJ1
and MPD1; MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1 and
DER1; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and HAC1;
MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1; MDJ2
and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2 and
HRD3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and HAC1; ERO1 and ERV2;
ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2; ERO1 and EPS1; ERO1
and PDI1; ERO1 and DER1; ERO1 and HRD3; ERO1 and UBC7; ERO1 and
DOA4; ERO1 and HAC1; ERV2 and EUG1; ERV2 and MPD1; ERV2 and MPD2;
ERV2 and EPS1; ERV2 and PDI1; ERV2 and DER1; ERV2 and HRD3; ERV2
and UBC7; ERV2 and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1 and
MPD2; EUG1 and EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and HRD3;
EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2; MPD1
and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and HRD3; MPD1 and
UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1; MPD2 and PDI1;
MPD2 and DER1; MPD2 and HRD3; MPD2 and UBC7; MPD2 and DOA4; MPD2
and HAC1; EPS1 and PDI1; EPS1 and DER1; EPS1 and HRD3; EPS1 and
UBC7; EPS1 and DOA4; EPS1 and HAC1; PDI1 and DER1; PDI1 and HRD3;
PDI1 and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and HRD3; DER1
and UBC7; DER1 and DOA4; DER1 and HAC1; HRD3 and UBC7; HRD3 and
DOA4; HRD3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0273] HRD3 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1
and SSB2; JEM1 and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and
ERO1; JEM1 and ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2;
JEM1 and EPS1; JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1
and UBC7; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1 LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1
and SSB1; LHS1 and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and
MDJ2; LHS1 and ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1;
LHS1 and MPD2; LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1
and DER3; LHS1 and UBC7; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1
and SSB1; SCJ1 and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and
MDJ2; SCJ1 and ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1;
SCJ1 and MPD2; SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1
and DER3; SCJ1 and UBC7; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSB2; KAR2 and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and
ERO1; KAR2 and ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2;
KAR2 and EPS1; KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2
and UBC7; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1;
SIL1 and SSE2; SEM and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1
and MDJ1; SIL1 and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and
EUG1; SIL1 and MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1;
SIL1 and DER1; SIL1 and DER3; SIL1 and UBC7; SIL1 and DOA4; SIL1
and HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and
SSA4; FKB2 and SSE1; FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2;
FKB2 and ECM10; FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2
and ERV2; FKB2 and EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and
EPS1; FKB2 and PDI1; FKB2 and DER1; FKB2 and DER3; FKB2 and UBC7;
FKB2 and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1
and SSA4; SSA1 and SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and
SSB2; to SSA1 and ECM10; SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and
ERO1; SSA1 and ERV2; SSA1 and EUG1; SSA1 and MPD1; SSA1 and MPD2;
SSA1 and EPS1; SSA1 and PDI1; SSA1 and DER1; SSA1 and DER3; SSA1
and UBC7; SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and
SSA4; SSA2 and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2;
SSA2 and ECM10; SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2
and ERV2; SSA2 and EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and
EPS1; SSA2 and PDI1; SSA2 and DER1; SSA2 and DER3; SSA2 and UBC7;
SSA2 and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3
and SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3 and
MDJ1; SSA3 and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and EUG1;
SSA3 and MPD1; SSA3 and MPD2; SSA3 and EPS1; SSA3 and PDI1; SSA3
and DER1; SSA3 and DER3; SSA3 and UBC7; SSA3 and DOA4; SSA3 and
HAC1; SSA4 and SSE1; SSA4 and SSE2; SSA4 and SSB1; SSA4 and SSB2;
SSA4 and ECM10; SSA4 and MDJ1; SSA4 and MDJ2; SSA4 and ERO1; SSA4
and ERV2; SSA4 and EUG1; SSA4 and MPD1; SSA4 and MPD2; SSA4 and
EPS1; SSA4 and PDI1; SSA4 and DER1; SSA4 and DER3; SSA4 and UBC7;
SSA4 and DOA4; SSA4 and HAC1; SSE1 and SSE2; SSE1 and SSB1; SSE1
and SSB2; SSE1 and ECM10; SSE1 and MDJ1; SSE1 and MDJ2; SSE1 and
ERO1; SSE1 and ERV2; SSE1 and EUG1; SSE1 and MPD1; SSE1 and MPD2;
SSE1 and EPS1; SSE1 and PDI1; SSE1 and DER1; SSE1 and DER3; SSE1
and UBC7; SSE1 and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and
SSB2; SSE2 and ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1;
SSE2 and ERV2; SSE2 and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2
and EPS1; SSE2 and PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and
UBC7; SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10;
SSB1 and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1
and EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and
PDI1; SSB1 and DER1; SSB1 and DER3; SSB1 and UBC7; SSB1 and DOA4;
SSB1 and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2 and MDJ2; SSB2
and ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and MPD1; SSB2 and
MPD2; SSB2 and EPS1; SSB2 and PDI1; SSB2 and DER1; SSB2 and DER3;
SSB2 and UBC7; SSB2 and DOA4; SSB2 and HAC1; ECM10 and MDJ1; ECM10
and MDJ2; ECM10 and ERO1; ECM10 and ERV2; ECM10 and EUG1; ECM10 and
MPD1; ECM10 and MPD2; ECM10 and EPS1; ECM10 and PDI1; ECM10 and
DER1; ECM10 and DER3; ECM10 and UBC7; ECM10 and DOA4; ECM10 and
HAC1; MDJ1 and MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1;
MDJ1 and MPD1; MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1
and DER1; MDJ1 and DER3; MDJ1 and UBC7; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2
and DER3; MDJ2 and UBC7; MDJ2 and DOA4; MDJ2 and HAC1; ERO1 and
ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2; ERO1 and EPS1;
ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1 and UBC7; ERO1
and DOA4; ERO1 and HAC1; ERV2 and EUG1; ERV2 and MPD1; ERV2 and
MPD2; ERV2 and EPS1; ERV2 and PDI1; ERV2 and DER1; ERV2 and DER3;
ERV2 and UBC7; ERV2 and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1
and MPD2; EUG1 and EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and
DER3; EUG1 and UBC7; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2;
MPD1 and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1
and UBC7; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1; MPD2 and
PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and UBC7; MPD2 and DOA4;
MPD2 and HAC1; EPS1 and PDI1; EPS1 and DER1; EPS1 and DER3; EPS1
and UBC7; EPS1 and DOA4; EPS1 and HAC1; PDI1 and DER1; PDI1 and
DER3; PDI1 and UBC7; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3;
DER1 and UBC7; DER1 and DOA4; DER1 and HAC1; DER3 and UBC7; DER3
and DOA4; DER3 and HAC1; UBC7 and DOA4; UBC7 and HAC1; or DOA4 and
HAC1.
[0274] UBC7 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1
and SSB2; JEM1 and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and
ERO1; JEM1 and ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2;
JEM1 and EPS1; JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1
and HRD3; JEM1 and DOA4; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1 LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1
and SSB1; LHS1 and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and
MDJ2; LHS1 and ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1;
LHS1 and MPD2; LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1
and DER3; LHS1 and HRD3; LHS1 and DOA4; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1
and SSB1; SCJ1 and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and
MDJ2; SCJ1 and ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1;
SCJ1 and MPD2; SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1
and DER3; SCJ1 and HRD3; SCJ1 and DOA4; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSB2; KAR2 and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and
ERO1; KAR2 and ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2;
KAR2 and EPS1; KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2
and HRD3; KAR2 and DOA4; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1;
SIL1 and SSE2; SIL1 and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1
and MDJ1; SIL1 and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and
EUG1; SIL1 and MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1;
SIL1 and DER1; SIL1 and DER3; SIL1 and HRD3; SIL1 and DOA4; SIL1
and HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and
SSA4; FKB2 and SSE1; FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2;
FKB2 and ECM10; FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2
and ERV2; FKB2 and EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and
EPS1; FKB2 and PDI1; FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3;
FKB2 and DOA4; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1
and SSA4; SSA1 and SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and
SSB2; SSA1 and ECM10; SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1;
SSA1 and ERV2; SSA1 and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1
and EPS1; SSA1 and PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and
HRD3; SSA1 and DOA4; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4;
SSA2 and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2
and ECM10; SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and
ERV2; SSA2 and EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1;
SSA2 and PDI1; SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2
and DOA4; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3 and
SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3 and MDJ1;
SSA3 and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and EUG1; SSA3
and MPD1; SSA3 and MPD2; SSA3 and EPS1; SSA3 and PDI1; SSA3 and
DER1; SSA3 and DER3; SSA3 and HRD3; SSA3 and DOA4; SSA3 and HAC1;
SSA4 and SSE1; SSA4 and SSE2; SSA4 and SSB1; SSA4 and SSB2; SSA4
and ECM10; SSA4 and MDJ1; SSA4 and MDJ2; SSA4 and ERO1; SSA4 and
ERV2; SSA4 and EUG1; SSA4 and MPD1; SSA4 and MPD2; SSA4 and EPS1;
SSA4 and PDI1; SSA4 and DER1; SSA4 and DER3; SSA4 and HRD3; SSA4
and DOA4; SSA4 and HAC1; SSE1 and SSE2; SSE1 and SSB1; SSE1 and
SSB2; SSE1 and ECM10; SSE1 and MDJ1; SSE1 and MDJ2; SSE1 and ERO1;
SSE1 and ERV2; SSE1 and EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1
and EPS1; SSE1 and PDI1; SSE1 and DER1; SSE1 and DER3; SSE1 and
HRD3; SSE1 and DOA4; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2;
SSE2 and ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2
and ERV2; SSE2 and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and
EPS1; SSE2 and PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3;
SSE2 and DOA4; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10; SSB1
and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1 and
EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and PDI1;
SSB1 and DER1; SSB1 and DER3; SSB1 and HRD3; SSB1 and DOA4; SSB1
and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2 and MDJ2; SSB2 and
ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and MPD1; SSB2 and MPD2;
SSB2 and EPS1; SSB2 and PDI1; SSB2 and DER1; SSB2 and DER3; SSB2
and HRD3; SSB2 and DOA4; SSB2 and HAC1; ECM10 and MDJ1; ECM10 and
MDJ2; ECM10 and ERO1; ECM10 and ERV2; ECM10 and EUG1; ECM10 and
MPD1; ECM10 and MPD2; ECM10 and EPS1; ECM10 and PDI1; ECM10 and
DER1; ECM10 and DER3; ECM10 and HRD3; ECM10 and DOA4; ECM10 and
HAC1; MDJ1 and MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1;
MDJ1 and MPD1; MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1
and DER1; MDJ1 and DER3; MDJ1 and HRD3; MDJ1 and DOA4; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2
and DER3; MDJ2 and HRD3; MDJ2 and DOA4; MDJ2 and HAC1; ERO1 and
ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2; ERO1 and EPS1;
ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1 and HRD3; ERO1
and DOA4; ERO1 and HAC1; ERV2 and EUG1; ERV2 and MPD1; ERV2 and
MPD2; ERV2 and EPS1; ERV2 and PDI1; ERV2 and DER1; ERV2 and DER3;
ERV2 and HRD3; ERV2 and DOA4; ERV2 and HAC1; EUG1 and MPD1; EUG1
and MPD2; EUG1 and EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and
DER3; EUG1 and HRD3; EUG1 and DOA4; EUG1 and HAC1; MPD1 and MPD2;
MPD1 and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1
and HRD3; MPD1 and DOA4; MPD1 and HAC1; MPD2 and EPS1; MPD2 and
PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2 and DOA4;
MPD2 and HAC1; EPS1 and PDI1; EPS1 and DER1; EPS1 and DER3; EPS1
and HRD3; EPS1 and DOA4; EPS1 and HAC1; PDI1 and DER1; PDI1 and
DER3; PDI1 and HRD3; PDI1 and DOA4; PDI1 and HAC1; DER1 and DER3;
DER1 and HRD3; DER1 and DOA4; DER1 and HAC1; DER3 and HRD3; DER3
and DOA4; DER3 and HAC1; HRD3 and DOA4; HRD3 and HAC1; or DOA4 and
HAC1.
[0275] DOA4 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SIL1; JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1
and SSB2; JEM1 and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and
ERO1; JEM1 and ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2;
JEM1 and EPS1; JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1
and HRD3; JEM1 and UBC7; JEM1 and HAC1; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1; LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1
and SSB1; LHS1 and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and
MDJ2; LHS1 and ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1;
LHS1 and MPD2; LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1
and DER3; LHS1 and HRD3; LHS1 and UBC7; LHS1 and HAC1; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1
and SSB1; SCJ1 and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and
MDJ2; SCJ1 and ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1;
SCJ1 and MPD2; SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1
and DER3; SCJ1 and HRD3; SCJ1 and UBC7; SCJ1 and HAC1; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSB2; KAR2 and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and
ERO1; KAR2 and ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2;
KAR2 and EPS1; KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2
and HRD3; KAR2 and UBC7; KAR2 and HAC1; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1;
SIL1 and SSE2; SIL1 and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1
and MDJ1; SIL1 and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and
EUG1; SIL1 and MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1;
SIL1 and DER1; SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1
and HAC1; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and
SSA4; FKB2 and SSE1; FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2;
FKB2 and ECM10; FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2
and ERV2; FKB2 and EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and
EPS1; FKB2 and PDI1; FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3;
FKB2 and UBC7; FKB2 and HAC1; SSA1 and SSA2; SSA1 and SSA3; SSA1
and SSA4; SSA1 and SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and
SSB2; SSA1 and ECM10; SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1;
SSA1 and ERV2; SSA1 and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1
and EPS1; SSA1 and PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and
HRD3; SSA1 and UBC7; SSA1 and HAC1; SSA2 and SSA3; SSA2 and SSA4;
SSA2 and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2
and ECM10; SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and
ERV2; SSA2 and EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1;
SSA2 and PDI1; SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2
and UBC7; SSA2 and HAC1; SSA3 and SSA4; SSA3 and SSE1; SSA3 and
SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3 and MDJ1;
SSA3 and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and EUG1; SSA3
and MPD1; SSA3 and MPD2; SSA3 and EPS1; SSA3 and PDI1; SSA3 and
DER1; SSA3 and DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3 and HAC1;
SSA4 and SSE1; SSA4 and SSE2; SSA4 and SSB1; SSA4 and SSB2; SSA4
and ECM10; SSA4 and MDJ1; SSA4 and MDJ2; SSA4 and ERO1; SSA4 and
ERV2; SSA4 and EUG1; SSA4 and MPD1; SSA4 and MPD2; SSA4 and EPS1;
SSA4 and PDI1; SSA4 and DER1; SSA4 and DER3; SSA4 and HRD3; SSA4
and UBC7; SSA4 and HAC1; SSE1 and SSE2; SSE1 and SSB1; SSE1 and
SSB2; SSE1 and ECM10; SSE1 and MDJ1; SSE1 and MDJ2; SSE1 and ERO1;
SSE1 and ERV2; SSE1 and EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1
and EPS1; SSE1 and PDI1; SSE1 and DER1; SSE1 and DER3; SSE1 and
HRD3; SSE1 and UBC7; SSE1 and HAC1; SSE2 and SSB1; SSE2 and SSB2;
SSE2 and ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2
and ERV2; SSE2 and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and
EPS1; SSE2 and PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3;
SSE2 and UBC7; SSE2 and HAC1; SSB1 and SSB2; SSB1 and ECM10; SSB1
and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1 and
EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and PDI1;
SSB1 and DER1; SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7; SSB1
and HAC1; SSB2 and ECM10; SSB2 and MDJ1; SSB2 and MDJ2; SSB2 and
ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and MPD1; SSB2 and MPD2;
SSB2 and EPS1; SSB2 and PDI1; SSB2 and DER1; SSB2 and DER3; SSB2
and HRD3; SSB2 and UBC7; SSB2 and HAC1; ECM10 and MDJ1; ECM10 and
MDJ2; ECM10 and ERO1; ECM10 and ERV2; ECM10 and EUG1; ECM10 and
MPD1; ECM10 and MPD2; ECM10 and EPS1; ECM10 and PDI1; ECM10 and
DER1; ECM10 and DER3; ECM10 and HRD3; ECM10 and UBC7; ECM10 and
HAC1; MDJ1 and MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1;
MDJ1 and MPD1; MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1
and DER1; MDJ1 and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and
HAC1; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2
and DER3; MDJ2 and HRD3; MDJ2 and UBC7; MDJ2 and HAC1; ERO1 and
ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2; ERO1 and EPS1;
ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1 and HRD3; ERO1
and UBC7; ERO1 and HAC1; ERV2 and EUG1; ERV2 and MPD1; ERV2 and
MPD2; ERV2 and EPS1; ERV2 and PDI1; ERV2 and DER1; ERV2 and DER3;
ERV2 and HRD3; ERV2 and UBC7; ERV2 and HAC1; EUG1 and MPD1; EUG1
and MPD2; EUG1 and EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and
DER3; EUG1 and HRD3; EUG1 and UBC7; EUG1 and HAC1; MPD1 and MPD2;
MPD1 and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1
and HRD3; MPD1 and UBC7; MPD1 and HAC1; MPD2 and EPS1; MPD2 and
PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2 and UBC7;
MPD2 and HAC1; EPS1 and PDI1; EPS1 and DER1; EPS1 and DER3; EPS1
and HRD3; EPS1 and UBC7; EPS1 and HAC1; PDI1 and DER1; PDI1 and
DER3; PDI1 and HRD3; PDI1 and UBC7; PDI1 and HAC1; DER1 and DER3;
DER1 and HRD3; DER1 and UBC7; DER1 and HAC1; DER3 and HRD3; DER3
and UBC7; DER3 and HAC1; HRD3 and UBC7; HRD3 and HAC1; or UBC7 and
HAC1.
[0276] HAC1 in combination with any one of the following
combinations: JEM1 and LHS1; JEM1 and SCJ1; JEM1 and KAR2; JEM1 and
SELL JEM1 and FKB2; JEM1 and SSA1; JEM1 and SSA2; JEM1 and SSA3;
JEM1 and SSA4; JEM1 and SSE1; JEM1 and SSE2; JEM1 and SSB1; JEM1
and SSB2; JEM1 and ECM10; JEM1 and MDJ1; JEM1 and MDJ2; JEM1 and
ERO1; JEM1 and ERV2; JEM1 and EUG1; JEM1 and MPD1; JEM1 and MPD2;
JEM1 and EPS1; JEM1 and PDI1; JEM1 and DER1; JEM1 and DER3; JEM1
and HRD3; JEM1 and UBC7; JEM1 and DOA4; LHS1 and SCJ1; LHS1 and
KAR2; LHS1 and SIL1 LHS1 and FKB2; LHS1 and SSA1; LHS1 and SSA2;
LHS1 and SSA3; LHS1 and SSA4; LHS1 and SSE1; LHS1 and SSE2; LHS1
and SSB1; LHS1 and SSB2; LHS1 and ECM10; LHS1 and MDJ1; LHS1 and
MDJ2; LHS1 and ERO1; LHS1 and ERV2; LHS1 and EUG1; LHS1 and MPD1;
LHS1 and MPD2; LHS1 and EPS1; LHS1 and PDI1; LHS1 and DER1; LHS1
and DER3; LHS1 and HRD3; LHS1 and UBC7; LHS1 and DOA4; SCJ1 and
KAR2; SCJ1 and SIL1; SCJ1 and FKB2; SCJ1 and SSA1; SCJ1 and SSA2;
SCJ1 and SSA3; SCJ1 and SSA4; SCJ1 and SSE1; SCJ1 and SSE2; SCJ1
and SSB1; SCJ1 and SSB2; SCJ1 and ECM10; SCJ1 and MDJ1; SCJ1 and
MDJ2; SCJ1 and ERO1; SCJ1 and ERV2; SCJ1 and EUG1; SCJ1 and MPD1;
SCJ1 and MPD2; SCJ1 and EPS1; SCJ1 and PDI1; SCJ1 and DER1; SCJ1
and DER3; SCJ1 and HRD3; SCJ1 and UBC7; SCJ1 and DOA4; KAR2 and
SIL1; KAR2 and FKB2; KAR2 and SSA1; KAR2 and SSA2; KAR2 and SSA3;
KAR2 and SSA4; KAR2 and SSE1; KAR2 and SSE2; KAR2 and SSB1; KAR2
and SSB2; KAR2 and ECM10; KAR2 and MDJ1; KAR2 and MDJ2; KAR2 and
ERO1; KAR2 and ERV2; KAR2 and EUG1; KAR2 and MPD1; KAR2 and MPD2;
KAR2 and EPS1; KAR2 and PDI1; KAR2 and DER1; KAR2 and DER3; KAR2
and HRD3; KAR2 and UBC7; KAR2 and DOA4; SIL1 and FKB2; SIL1 and
SSA1; SIL1 and SSA2; SIL1 and SSA3; SIL1 and SSA4; SIL1 and SSE1;
SIL1 and SSE2; SIL1 and SSB1; SIL1 and SSB2; SIL1 and ECM10; SIL1
and MDJ1; SIL1 and MDJ2; SIL1 and ERO1; SIL1 and ERV2; SIL1 and
EUG1; SIL1 and MPD1; SIL1 and MPD2; SIL1 and EPS1; SIL1 and PDI1;
SIL1 and DER1; SIL1 and DER3; SIL1 and HRD3; SIL1 and UBC7; SIL1
and DOA4; FKB2 and SSA1; FKB2 and SSA2; FKB2 and SSA3; FKB2 and
SSA4; FKB2 and SSE1; FKB2 and SSE2; FKB2 and SSB1; FKB2 and SSB2;
FKB2 and ECM10; FKB2 and MDJ1; FKB2 and MDJ2; FKB2 and ERO1; FKB2
and ERV2; FKB2 and EUG1; FKB2 and MPD1; FKB2 and MPD2; FKB2 and
EPS1; FKB2 and PDI1; FKB2 and DER1; FKB2 and DER3; FKB2 and HRD3;
FKB2 and UBC7; FKB2 and DOA4; SSA1 and SSA2; SSA1 and SSA3; SSA1
and SSA4; SSA1 and SSE1; SSA1 and SSE2; SSA1 and SSB1; SSA1 and
SSB2; SSA1 and ECM10; SSA1 and MDJ1; SSA1 and MDJ2; SSA1 and ERO1;
SSA1 and ERV2; SSA1 and EUG1; SSA1 and MPD1; SSA1 and MPD2; SSA1
and EPS1; SSA1 and PDI1; SSA1 and DER1; SSA1 and DER3; SSA1 and
HRD3; SSA1 and UBC7; SSA1 and DOA4; SSA2 and SSA3; SSA2 and SSA4;
SSA2 and SSE1; SSA2 and SSE2; SSA2 and SSB1; SSA2 and SSB2; SSA2
and ECM10; SSA2 and MDJ1; SSA2 and MDJ2; SSA2 and ERO1; SSA2 and
ERV2; SSA2 and EUG1; SSA2 and MPD1; SSA2 and MPD2; SSA2 and EPS1;
SSA2 and PDI1; SSA2 and DER1; SSA2 and DER3; SSA2 and HRD3; SSA2
and UBC7; SSA2 and DOA4; SSA3 and SSA4; SSA3 and SSE1; SSA3 and
SSE2; SSA3 and SSB1; SSA3 and SSB2; SSA3 and ECM10; SSA3 and MDJ1;
SSA3 and MDJ2; SSA3 and ERO1; SSA3 and ERV2; SSA3 and EUG1; SSA3
and MPD1; SSA3 and MPD2; SSA3 and EPS1; SSA3 and PDI1; SSA3 and
DER1; SSA3 and DER3; SSA3 and HRD3; SSA3 and UBC7; SSA3 and DOA4;
SSA4 and SSE1; SSA4 and SSE2; SSA4 and SSB1; SSA4 and SSB2; SSA4
and ECM10; SSA4 and MDJ1; SSA4 and MDJ2; SSA4 and ERO1; SSA4 and
ERV2; SSA4 and EUG1; SSA4 and MPD1; SSA4 and MPD2; SSA4 and EPS1;
SSA4 and PDI1; SSA4 and DER1; SSA4 and DER3; SSA4 and HRD3; SSA4
and UBC7; SSA4 and DOA4; SSE1 and SSE2; SSE1 and SSB1; SSE1 and
SSB2; SSE1 and ECM10; SSE1 and MDJ1; SSE1 and MDJ2; SSE1 and ERO1;
SSE1 and ERV2; SSE1 and EUG1; SSE1 and MPD1; SSE1 and MPD2; SSE1
and EPS1; SSE1 and PDI1; SSE1 and DER1; SSE1 and DER3; SSE1 and
HRD3; SSE1 and UBC7; SSE1 and DOA4; SSE2 and SSB1; SSE2 and SSB2;
SSE2 and ECM10; SSE2 and MDJ1; SSE2 and MDJ2; SSE2 and ERO1; SSE2
and ERV2; SSE2 and EUG1; SSE2 and MPD1; SSE2 and MPD2; SSE2 and
EPS1; SSE2 and PDI1; SSE2 and DER1; SSE2 and DER3; SSE2 and HRD3;
SSE2 and UBC7; SSE2 and DOA4; SSB1 and SSB2; SSB1 and ECM10; SSB1
and MDJ1; SSB1 and MDJ2; SSB1 and ERO1; SSB1 and ERV2; SSB1 and
EUG1; SSB1 and MPD1; SSB1 and MPD2; SSB1 and EPS1; SSB1 and PDI1;
SSB1 and DER1; SSB1 and DER3; SSB1 and HRD3; SSB1 and UBC7; SSB1
and DOA4; SSB2 and ECM10; SSB2 and MDJ1; SSB2 and MDJ2; SSB2 and
ERO1; SSB2 and ERV2; SSB2 and EUG1; SSB2 and MPD1; SSB2 and MPD2;
SSB2 and EPS1; SSB2 and PDI1; SSB2 and DER1; SSB2 and DER3; SSB2
and HRD3; SSB2 and UBC7; SSB2 and DOA4; ECM10 and MDJ1; ECM10 and
MDJ2; ECM10 and ERO1; ECM10 and ERV2; ECM10 and EUG1; ECM10 and
MPD1; ECM10 and MPD2; ECM10 and EPS1; ECM10 and PDI1; ECM10 and
DER1; ECM10 and DER3; ECM10 and HRD3; ECM10 and UBC7; ECM10 and
DOA4; MDJ1 and MDJ2; MDJ1 and ERO1; MDJ1 and ERV2; MDJ1 and EUG1;
MDJ1 and MPD1; MDJ1 and MPD2; MDJ1 and EPS1; MDJ1 and PDI1; MDJ1
and DER1; MDJ1 and DER3; MDJ1 and HRD3; MDJ1 and UBC7; MDJ1 and
DOA4; MDJ2 and ERO1; MDJ2 and ERV2; MDJ2 and EUG1; MDJ2 and MPD1;
MDJ2 and MPD2; MDJ2 and EPS1; MDJ2 and PDI1; MDJ2 and DER1; MDJ2
and DER3; MDJ2 and HRD3; MDJ2 and UBC7; MDJ2 and DOA4; ERO1 and
ERV2; ERO1 and EUG1; ERO1 and MPD1; ERO1 and MPD2; ERO1 and EPS1;
ERO1 and PDI1; ERO1 and DER1; ERO1 and DER3; ERO1 and HRD3; ERO1
and UBC7; ERO1 and DOA4; ERV2 and EUG1; ERV2 and MPD1; ERV2 and
MPD2; ERV2 and EPS1; ERV2 and PDI1; ERV2 and DER1; ERV2 and DER3;
ERV2 and HRD3; ERV2 and UBC7; ERV2 and DOA4; EUG1 and MPD1; EUG1
and MPD2; EUG1 and EPS1; EUG1 and PDI1; EUG1 and DER1; EUG1 and
DER3; EUG1 and HRD3; EUG1 and UBC7; EUG1 and DOA4; MPD1 and MPD2;
MPD1 and EPS1; MPD1 and PDI1; MPD1 and DER1; MPD1 and DER3; MPD1
and HRD3; MPD1 and UBC7; MPD1 and DOA4; MPD2 and EPS1; MPD2 and
PDI1; MPD2 and DER1; MPD2 and DER3; MPD2 and HRD3; MPD2 and UBC7;
MPD2 and DOA4; EPS1 and PDI1; EPS1 and DER1; EPS1 and DER3; EPS1
and HRD3; EPS1 and UBC7; EPS1 and DOA4; PDI1 and DER1; PDI1 and
DER3; PDI1 and HRD3; PDI1 and UBC7; PDI1 and DOA4; DER1 and DER3;
DER1 and HRD3; DER1 and UBC7; DER1 and DOA4; DER3 and HRD3; DER3
and UBC7; DER3 and DOA4; HRD3 and UBC7; HRD3 and DOA4; or UBC7 and
DOA4.
Protein Product of Choice
[0277] In principle, any protein can be expressed as the protein
product of choice.
[0278] As discussed above, the protein product of choice may or may
not be a protein that is naturally produced by the host cell, in
which case the protein may or may not be encoded by the host cell's
endogenous gene for that protein or the protein may or may not be
encoded (fully, or in part) by an exogenous polynucleotide
sequence.
[0279] Thus, it is possible to produce enhanced levels of naturally
produced proteins by transforming the host cell with a
polynucleotide encoding a further, or replacement, copy of an
endogenous gene, or otherwise genetically modifying the host cell
to increase the expression of a naturally produced protein. In one
embodiment, a recombinant or genetically modified endogenous gene
has a sequence that is different to the endogenous genetic material
of the host cell.
[0280] The protein product of choice may or may not be a
heterologous protein, by which we mean that the protein is one that
is not naturally produced by the host cell. In the case of a
heterologous protein product of choice, the protein may or may not
be encoded by an exogenous polynucleotide sequence.
[0281] In one embodiment, the protein product of choice is
secreted. In that case, a sequence encoding a secretion leader
sequence which, for example, comprises most of the natural HSA
secretion leader, plus a small portion of the S. cerevisiae
.alpha.-mating factor secretion leader as taught in WO 90/01063 may
or may not be included in the open reading frame that encodes the
protein product of choice.
[0282] Alternatively, the protein product of choice may or may not
be intracellular.
[0283] It is known in the prior art that enhanced protein
production can be achieved by co-expression of a protein product
and a chaperone in different compartments of the cell. For example,
WO 2005/061718 (Example 12) describes the co-over-expression of the
cytoplasmic chaperone SSA1 and a secreted recombinant transferrin,
in order to increase the production of the secreted recombinant
transferrin.
[0284] In another preferred embodiment, the protein product of
choice comprises the sequence of a eukaryotic protein, or a
fragment or variant thereof. Suitable eukaryotes include fungi,
plants and animals. In one preferred embodiment the protein product
of choice is a fungal protein, such as a yeast protein. In another
preferred embodiment the protein product of choice is an animal
protein. Exemplary animals include vertebrates and invertebrates.
Exemplary vertebrates include mammals, such as humans, and
non-human mammals.
[0285] Thus the protein product of choice may or may not comprise
the sequence of a yeast protein.
[0286] The protein product of choice may or may not comprise
albumin, a monoclonal antibody, an etoposide, a serum protein (such
as a blood clotting factor), antistasin, a tick anticoagulant
peptide, transferrin, lactoferrin, endostatin, angiostatin,
collagens, immunoglobulins or immunoglobulin-based-molecules or
fragment of either (e.g. a Small Modular ImmunoPharmaceutical.TM.
("SMTP") or dAb, Fab' fragments, F(ab').sub.2, scAb, scFv or scFv
fragment), a Kunitz domain protein (such as aprotinin, amyloid
precursor protein and those described in WO 03/066824, with or
without albumin fusions), interferons (such as interferon .alpha.
species and sub-species, interferon (3 species and sub-species,
interferon .gamma. species and sub-species), interleukins (such as
IL10, IL11 and IL2), leptin, CNTF and fragment thereof (such as
CNTF.sub.Ax15 (Axokine.TM.)), IL1-receptor antagonist,
erythropoietin (EPO) and EPO mimics, thrombopoietin (TPO) and TPO
mimics, prosaptide, cyanovirin-N, 5-helix, T20 peptide, T1249
peptide, HIV gp41, HIV gp120, urokinase, prourokinase, tPA,
hirudin, platelet derived growth factor, parathyroid hormone,
proinsulin, insulin, glucagon, glucagon-like peptides, insulin-like
growth factor, calcitonin, growth hormone, transforming growth
factor .beta., tumour necrosis factor, G-CSF, GM-CSF, M-CSF, FGF,
coagulation factors in both pre and active forms, including but not
limited to plasminogen, fibrinogen, thrombin, pre-thrombin,
pro-thrombin, von Willebrand's factor, .alpha..sub.1-antitrypsin,
plasminogen activators, Factor VII, Factor Vac Factor IX, Factor X
and Factor Mil, nerve growth factor, LACI, platelet-derived
endothelial cell growth factor (PD-ECGF), glucose oxidase, serum
cholinesterase, inter-alpha trypsin inhibitor, antithrombin III,
apo-lipoprotein species, Protein C, Protein S, or a variant or
fragment of any of the above.
[0287] A "variant", in the context of the above-listed proteins,
refers to a protein wherein at one or more positions there have
been amino acid insertions, deletions, or substitutions, either
conservative or non-conservative, provided that such changes result
in a protein whose basic properties, for example enzymatic activity
or receptor binding (type of and specific activity),
thermostability, activity in a certain pH-range (pH-stability) have
not significantly been changed. "Significantly" in this context
means that one skilled in the art would say that the properties of
the variant may still be different but would not be unobvious over
the ones of the original protein.
[0288] By "conservative substitutions" is intended combinations
such as Val, Ile, Leu, Ala, Met; Asp, Glu; Asn, Gln; Ser, Thr, Gly,
Ala; Lys, Arg, His; and Phe, Tyr, Trp. Preferred conservative
substitutions include Gly, Ala; Val, Ile, Leu; Asp, Glu; Asn, Gln;
Ser, Thr; Lys, Arg; and Phe, Tyr.
[0289] A "variant" typically has at least 25%, at least 50%, at
least 60% or at least 70%, preferably at least 80%, more preferably
at least 90%, even more preferably at least 95%, yet more
preferably at least 99%, most preferably at least 99.5% sequence
identity to the polypeptide from which it is derived.
[0290] The percent sequence identity between two polypeptides may
be determined using suitable computer programs, for example the GAP
program of the University of Wisconsin Genetic Computing Group and
it will be appreciated that percent identity is calculated in
relation to polypeptides whose sequence has been aligned
optimally.
[0291] The alignment may alternatively be carried out using the
Clustal W program (Thompson et al., (1994) Nucleic Acids Res.,
22(22), 4673-80). The parameters used may be as follows: [0292]
Fast pairwise alignment parameters: K-tuple(word) size; 1, window
size; 5, gap penalty; 3, number of top diagonals; 5. Scoring
method: x percent. [0293] Multiple alignment parameters: gap open
penalty; 10, gap extension penalty; 0.05. [0294] Scoring matrix:
BLOSUM.
[0295] Such variants may or may not be natural or made using the
methods of protein engineering and site-directed mutagenesis as are
well known in the art.
[0296] A "fragment", in the context of the above-listed proteins,
refers to a protein wherein at one or more positions there have
been deletions. Thus the fragment may comprise at most 5, 10, 20,
30, 40 or 50% of the complete sequence of the full mature
polypeptide. Typically a fragment comprises up to 60%, more
typically up to 70%, preferably up to 80%, more preferably up to
90%, even more preferably up to 95%, yet more preferably up to 99%
of the complete sequence of the full desired protein. Particularly
preferred fragments of a protein comprise one or more whole domains
of the protein.
[0297] In one particularly preferred embodiment the protein product
of choice comprises the sequence of albumin or a variant or
fragment thereof.
[0298] By "albumin" we include a protein comprising the sequence of
an albumin protein obtained from any source. Typically the source
is mammalian. In one preferred embodiment the serum albumin is
human serum albumin ("HSA"). The term "human serum albumin"
includes the meaning of a serum albumin having an amino acid
sequence naturally occurring in humans, and variants thereof.
Preferably the albumin has the amino acid sequence disclosed in WO
90/13653 or a variant thereof. The HSA coding sequence is
obtainable by known methods for isolating cDNA corresponding to
human genes, and is also disclosed in, for example, EP 73 646 and
EP 286 424.
[0299] In another preferred embodiment the "albumin" comprises the
sequence of bovine serum albumin. The term "bovine serum albumin"
includes the meaning of a serum albumin having an amino acid
sequence naturally occurring in cows, for example as taken from
Swissprot accession number P02769, and variants thereof as defined
below. The term "bovine serum albumin" also includes the meaning of
fragments, of full-length bovine serum albumin or variants thereof,
as defined below.
[0300] In another preferred embodiment the albumin comprises the
sequence of an albumin derived from one of serum albumin from dog
(e.g. see Swissprot accession number P49822), pig (e.g. see
Swissprot accession number P08835), goat (e.g. as available from
Sigma as product no. A2514 or A4164), turkey (e.g. see Swissprot
accession number O73860), baboon (e.g. as available from Sigma as
product no. A1516), cat (e.g. see Swissprot accession number
P49064), chicken (e.g. see Swissprot accession number P19121),
ovalbumin (e.g. chicken ovalbumin) (e.g. see Swissprot accession
number P01012), donkey (e.g. see Swissprot accession number
P39090), guinea pig (e.g. as available from Sigma as product no.
A3060, A2639, O5483 or A6539), hamster (e.g. as available from
Sigma as product no. A5409), horse (e.g. see Swissprot accession
number P35747), rhesus monkey (e.g. see Swissprot accession number
Q28522), mouse (e.g. see Swissprot accession number O89020), pigeon
(e.g. as defined by Khan et al, 2002, Int. J. Biol. Macromol.,
30(3-4)371-8), rabbit (e.g. see Swissprot accession number P49065),
rat (e.g. see Swissprot accession number P36953) and sheep (e.g.
see Swissprot accession number P14639) and includes variants and
fragments thereof as defined below.
[0301] Many naturally occurring mutant forms of albumin are known.
Many are described in Peters, (1996, All About Albumin:
Biochemistry, Genetics and Medical Applications, Academic Press,
Inc., San Diego, Calif., p. 170-181). A variant as defined above
may or may not be one of these naturally occurring mutants.
[0302] A "variant albumin" refers to an albumin protein wherein at
one or more positions there have been amino acid insertions,
deletions, or substitutions, either conservative or
non-conservative, provided that such changes result in an albumin
protein for which at least one basic property, for example binding
activity (type of and specific activity e.g. binding to bilirubin),
osmolarity (oncotic pressure, colloid osmotic pressure), behaviour
in a certain pH-range (pH-stability) has not significantly been
changed. "Significantly" in this context means that one skilled in
the art would say that the properties of the variant may still be
different but would not be unobvious over the ones of the original
protein.
[0303] By "conservative substitutions" is intended combinations
such as Gly, Ala; Val, Ile, Leu; Asp, Glu; Asn, Gln; Ser, Thr, Lys,
Arg; and Phe, Tyr. Such variants may be made by techniques well
known in the art, such as by site-directed mutagenesis as disclosed
in U.S. Pat. No. 4,302,386 issued 24 Nov. 1981 to Stevens,
incorporated herein by reference.
[0304] Typically an albumin variant will have more than 40%,
usually at least 50%, more typically at least 60%, preferably at
least 70%, more preferably at least 80%, yet more preferably at
least 90%, even more preferably at least 95%, most preferably at
least 98% or more sequence identity with naturally occurring
albumin. The percent sequence identity between two polypeptides may
be determined using suitable computer programs, for example the GAP
program of the University of Wisconsin Genetic Computing Group and
it will, be appreciated that percent identity is calculated in
relation to polypeptides whose sequence has been aligned optimally.
The alignment may alternatively be carried out using the Clustal W
program (Thompson et al., 1994). The parameters used may be as
follows:
[0305] Fast pairwise alignment parameters: K-tuple(word) size; 1,
window size; 5, gap penalty; 3, number of top diagonals; 5. Scoring
method: x percent. Multiple alignment parameters: gap open penalty;
10, gap extension penalty; 0.05. Scoring matrix: BLOSUM.
[0306] The term "fragment" as used above includes any fragment of
full-length albumin or a variant thereof, so long as at least one
basic property, for example binding activity (type of and specific
activity e.g. binding to bilirubin), osmolarity (oncotic pressure,
colloid osmotic pressure), behaviour in a certain pH-range
(pH-stability) has not significantly been changed. "Significantly"
in this context means that one skilled in the art would say that
the properties of the variant may still be different but would not
be unobvious over the ones of the original protein. A fragment will
typically be at least 50 amino acids long. A fragment may or may
not comprise at least one whole sub-domain of albumin Domains of
HSA have been expressed as recombinant proteins (Dockal, M. et al.,
1999, J. Biol. Chem., 274, 29303-29310), where domain I was defined
as consisting of amino acids 1-197, domain II was defined as
consisting of amino acids 189-385 and domain III was defined as
consisting of amino acids 381-585. Partial overlap of the domains
occurs because of the extended .alpha.-helix structure (h10-h1)
which exists between domains I and II, and between domains II and
III (Peters, 1996, op. cit., Table 2-4). HSA also comprises six
sub-domains (sub-domains IA, IB, IIA, IIB, IIIA and IIIB).
Sub-domain IA comprises amino acids 6-105, sub-domain IB comprises
amino acids 120-177, sub-domain IIA comprises amino acids 200-291,
sub-domain IIB comprises amino acids 316-369, sub-domain IIIA
comprises amino acids 392-491 and sub-domain IIIB comprises amino
acids 512-583. A fragment may or may not comprise a whole or part
of one or more domains or sub-domains as defined above, or any
combination of those domains and/or sub-domains.
[0307] In another particularly preferred embodiment the protein
product of choice comprises the sequence of transferrin or a
variant or fragment thereof. The term "transferrin" as used herein
includes all members of the transferrin family (Testa, Proteins of
iron metabolism, CRC Press, 2002; Harris & Aisen, Iron carriers
and iron proteins, Vol. 5, Physical Bioinorganic Chemistry, VCH,
1991) and their derivatives, such as transferrin, mutant
transferrins (Mason et al, 1993, Biochemistry, 32, 5472; Mason et
al, 1998, Biochem. J., 330(1), 35), truncated transferrins,
transferrin lobes (Mason et al, 1996, Protein Expr. Purif., 8, 119;
Mason et al, 1991, Protein Expr. Purif., 2, 214), lactoferrin,
mutant lactoferrins, truncated lactoferrins, lactoferrin lobes or
fusions of any of the above to other peptides, polypeptides or
proteins (Shin et al, 1995, Proc. Natl. Acad Sci. USA, 92, 2820;
Ali et al, 1999, J. Biol. Chem., 274, 24066; Mason et al, 2002,
Biochemistry, 41, 9448).
[0308] The transferrin may or may not be human transferrin. The
term "human transferrin" is used herein to denote material which is
indistinguishable from transferrin derived from a human or which is
a variant or fragment thereof. A "variant" includes insertions,
deletions and substitutions, either conservative or
non-conservative, where such changes do not substantially alter the
useful ligand-binding or immunogenic properties of transferrin.
[0309] Mutants of transferrin are included in the invention. Such
mutants may or may not have altered immunogenicity. For example,
transferrin mutants may or may not display modified (e.g. reduced)
glycosylation. The N-linked glycosylation pattern of a transferrin
molecule can be modified by adding/removing amino acid
glycosylation consensus sequences such as N-X-S/T, at any or all of
the N, X, or S/T position. Transferrin mutants may or may not be
altered in their natural binding to metal ions and/or other
proteins, such as transferrin receptor. An example of a transferrin
mutant modified in this manner is exemplified below.
[0310] We also include naturally-occurring polymorphic variants of
human transferrin or human transferrin analogues. Generally,
variants or fragments of human transferrin will have at least 5%,
10%, 15%, 20%, 30%, 40% or 50% (preferably at least 80%, 90% or
95%) of human transferrin's ligand binding activity (for example
iron-binding), weight for weight. The iron binding activity of
transferrin or a test sample can be determined
spectrophotometrically by 470 nm:280 nm absorbance ratios for the
proteins in their iron-free and fully iron-loaded states. Reagents
should be iron-free unless stated otherwise. Iron can be removed
from transferrin or the test sample by dialysis against 0.1M
citrate, 0.1M acetate, 10 mM EDTA pH4.5. Protein should be at
approximately 20 mg/mL in 100 mM HEPES, 10 mM NaHCO.sub.3 pH8.0.
Measure the 470 nm:280 nm absorbance ratio of apo-transferrin
(Calbiochem, CN Biosciences, Nottingham, UK) diluted in water so
that absorbance at 280 nm can be accurately determined
spectrophotometrically (0% iron binding). Prepare 20 mM
iron-nitrilotriacetate (FeNTA) solution by dissolving 191 mg
nitrotriacetic acid in 2 mT, 1M NaOH, then add 2 mL 0.5M ferric
chloride. Dilute to 50 mL with deionised water. Fully load
apo-transferrin with iron (100% iron binding) by adding a
sufficient excess of freshly prepared 20 mM FeNTA, then dialyse the
holo-transferrin preparation completely against 100 mM HEPES, 10 mM
NaHCO.sub.3 pH8.0 to remove remaining FeNTA before measuring the
absorbance ratio at 470 nm:280 nm. Repeat the procedure using test
sample, which should initially be free from iron, and compare final
ratios to the control. Additionally, single or multiple
heterologous fusions comprising any of the above; or single or
multiple heterologous fusions to albumin, transferrin or
immunoglobulins or a variant or fragment of any of these may be
used. Such fusions include albumin N-terminal fusions, albumin
C-terminal fusions and co-N-terminal and C-terminal albumin fusions
as exemplified by WO 01/79271, and transferrin N-terminal fusions,
transferrin C-terminal fusions, and co-N-terminal and C-terminal
transferrin fusions.
[0311] Examples of transferrin fusions are given in US patent
applications US2003/0221201 and US2003/0226155, Shin, et al., 1995,
Proc Natl Acad Sci USA, 92, 2820, Ali, et al., 1999, J Biol Chem,
274, 24066, Mason, et al., 2002, Biochemistry, 41, 9448, the
contents of which are incorporated herein by reference.
[0312] The skilled person will also appreciate that the open
reading frame of any other gene or variant, or part or either, can
be utilised as an open reading frame for use with the present
invention. For example, the open reading frame may encode a protein
comprising any sequence, be it a natural protein (including a
zymogen), or a variant, or a fragment (which may or may not, for
example, be a domain) of a natural protein; or a totally synthetic
protein; or a single or multiple fusion of different proteins
(natural or synthetic). Such proteins can be taken, but not
exclusively, from the lists provided in WO 01/79258, WO 01/79271,
WO 01/79442, WO 01/79443, WO 01/79444 and WO 01/79480, or a variant
or fragment thereof; the disclosures of which are incorporated
herein by reference. Although these patent applications present the
list of proteins in the context of fusion partners for albumin, the
present invention is not so limited and, for the purposes of the
present invention, any of the proteins listed therein may be
presented alone or as fusion partners for albumin, the Fc region of
immunoglobulin, transferrin, lactoferrin or any other protein or
fragment or variant of any of the above, as a desired
polypeptide.
[0313] The protein product of choice may or may not be a
therapeutically active protein. In other words, it may or may not
have a recognised medical effect on individuals, such as humans.
Many different types of therapeutically active protein are well
known in the art.
[0314] As discussed above, the protein product of choice may or may
not comprise a leader sequence effective to cause secretion in the
host cell (such as in a yeast host cell).
[0315] Numerous natural or artificial polypeptide signal sequences
(also called secretion pre regions) have been used or developed for
secreting proteins from host cells. The signal sequence directs the
nascent protein towards the machinery of the cell that exports
proteins from the cell into the surrounding medium or, in some
cases, into the periplasmic space. The signal sequence is usually,
although not necessarily, located at the N-terminus of the primary
translation product and is generally, although not necessarily,
cleaved off the protein during the secretion process, to yield the
"mature" protein.
[0316] In the case of some proteins the entity that is initially
secreted, after the removal of the signal sequence, includes
additional amino acids at its N-terminus called a "pro" sequence,
the intermediate entity being called a "pro-protein". These pro
sequences may assist the final protein to fold and become
functional, and are usually then cleaved off. In other instances,
the pro region simply provides a cleavage site for an enzyme to
cleave off the pre-pro region and is not known to have another
function.
[0317] The pro sequence can be removed either during the secretion
of the protein from the cell or after export from the cell into the
surrounding medium or periplasmic space.
[0318] Polypeptide sequences which direct the secretion of
proteins, whether they resemble signal (i.e. pre) sequences or
pre-pro secretion sequences, are referred to as leader sequences.
The secretion of proteins is a dynamic process involving
translation, translocation and post-translational processing, and
one or more of these steps may not necessarily be completed before
another is either initiated or completed.
[0319] For production of proteins in eukaryotic species such as the
yeasts Saccharomyces cerevisiae, Zygosaccharomyces species,
Kluyveromyces lactis and Pichia pastoris, known leader sequences
include those from the S. cerevisiae acid phosphatase protein
(Pho5p) (see EP 366 400), the invertase protein (Suc2p) (see Smith
et al. (1985) Science, 229, 1219-1224) and heat-shock protein-150
(Hsp150p) (see WO 95/33833). Additionally, leader sequences from
the S. cerevisiae mating factor alpha-1 protein (MF.alpha.-1) and
from the human lysozyme and human serum albumin (HSA) protein have
been used, the latter having been used especially, although not
exclusively, for secreting human albumin. WO 90/01063 discloses a
fusion of the MF.alpha.-1 and HSA leader sequences, which
advantageously reduces the production of a contaminating fragment
of human albumin relative to the use of the MF.alpha.-1 leader
sequence. Modified leader sequences are also disclosed in the
examples of this application and the reader will appreciate that
those leader sequences can be used with proteins other than
transferrin. In addition, the natural transferrin leader sequence
may or may not be used to direct secretion of transferrin and other
protein products of choice.
[0320] Where a helper protein is a chaperone involved in the
formation of disulphide bonds, then in one embodiment the protein
product of choice comprises disulphide bonds in its mature form.
The disulphide bonds may be intramolecular and/or
intermolecular.
[0321] The protein product of choice may or may not be a
commercially useful protein. Some heterologously expressed proteins
are intended to interact with the cell in which they are expressed
in order to bring about a beneficial effect on the cell's
activities. These proteins are not, in their own right,
commercially useful. Commercially useful proteins are proteins that
have a utility ex vivo of the cell in which they are expressed.
Nevertheless, the skilled reader will appreciate that a
commercially useful protein may or may not also have a biological
effect on the host cell expressing it as a protein, but that that
effect is not the main or sole reason for expressing the protein
therein.
Suitable Host Cells for the Practice of the Present Invention
[0322] The host cell may be any type of cell. The host cell may or
may not be an animal (such as mammalian, avian, insect, etc.),
plant, fungal or bacterial cell. Bacterial and fungal, such as
yeast, host cells may or may not be preferred.
[0323] Thus, the host cell may or may not be an animal (such as
mammalian, avian, insect, etc.) cell. Suitable methods for
transformation of animal cells are well known in the art and
include, for example the use of retrovirus vectors (such as
lentivirus vectors). Wolkowicz et al, 2004, Methods Mol. Biol.,
246, 391-411 describes the use of lentivirus vectors for delivery
of recombinant nucleic acid sequences to mammalian cells for use in
cell culture techniques. Fassler, 2004, EMBO Rep., 5(1), 28-9
reviews lentiviral transgene vectors and their use in the
production of transgenic systems.
[0324] In one embodiment the host cell is a yeast cell, such as a
member of the Saccharomyces, Kluyveromyces, or Pichia genus, such
as Saccharomyces cerevisiae, Kluyveromyces lactis, Pichia pastoris
and Pichia membranaefaciens, or Zygosaccharomyces rouxii,
Zygosaccharomyces bailii, Zygosaccharomyces fermentati, Hansenula
polymorpha (also known as Pichia angusta) or Kluyveromyces
drosophilarum are preferred.
[0325] It may be particularly advantageous to use a yeast deficient
in one or more protein mannosyl transferases involved in
O-glycosylation of proteins, for instance by disruption of the gene
coding sequence.
[0326] Recombinantly expressed proteins can be subject to
undesirable post-translational modifications by the producing host
cell. For example, the albumin protein sequence does not contain
any sites for N-linked glycosylation and has not been reported to
be modified, in nature, by O-linked glycosylation. However, it has
been found that recombinant human albumin ("rHA") produced in a
number of yeast species can be modified by O-linked glycosylation,
generally involving mannose. The mannosylated albumin is able to
bind to the lectin Concanavalin A. The amount of mannosylated
albumin produced by the yeast can be reduced by using a yeast
strain deficient in one or more of the PMT genes (WO 94/04687). The
most convenient way of achieving this is to create a yeast which
has a defect in its genome such that a reduced level of one of the
Pmt proteins is produced. For example, there may or may not be a
deletion, insertion or transposition in the coding sequence or the
regulatory regions (or in another gene regulating the expression of
one of the PMT genes) such that little or no Pmt protein is
produced. Alternatively, the yeast could be transformed to produce
an anti-Pmt agent, such as an anti-Pmt antibody. Alternatively, the
yeast could be cultured in the presence of a compound that inhibits
the activity of one of the PMT genes (Duffy et al, "Inhibition of
protein mannosyltransferase 1 (PMT1) activity in the pathogenic
yeast Candida albicans", International Conference on Molecular
Mechanisms of Fungal Cell Wall Biogenesis, 26-31 Aug. 2001, Monte
Verita, Switzerland, Poster Abstract P38; the poster abstract may
be viewed at http://www.micro.biol.ethz.ch/cellwall/).
[0327] If a yeast other than S. cerevisiae is used, disruption of
one or more of the genes equivalent to the PMT genes of S.
cerevisiae is also beneficial, e.g. in Pichia pastoris or
Kluyveromyces lactis. The sequence of PMT1 (or any other PMT gene)
isolated from S. cerevisiae may be used for the identification or
disruption of genes encoding similar enzymatic activities in other
fungal species. The cloning of the PMT1 homologue of Kluyveromyces
lactis is described in WO 94/04687.
[0328] The yeast may or may not also have a deletion of the HSP150
and/or YAP3 genes as taught respectively in WO 95/33833 and WO
95/23857.
[0329] Where one or more of the helper protein(s) and/or protein
product of choice are encoded by a plasmid-borne polynucleotide
sequence, the host cell type may be selected for compatibility with
the plasmid type being used.
[0330] The skilled person will appreciate that any suitable plasmid
may be used, such as a centromeric plasmid. The examples provide
suitable plasmids (centromeric YCplac33-based vectors) for use to
transform yeast host cells of the present invention. Alternatively,
any other suitable plasmid may be used, such as a yeast-compatible
2 .mu.m-based plasmid.
[0331] Plasmids obtained from one yeast type can be maintained in
other yeast types (Irie et al, 1991, Gene, 108(1), 139-144; Irie et
al, 1991, Mol. Gen. Genet., 225(2), 257-265). For example, pSR1
from Zygosaccharomyces rouxii can be maintained in Saccharomyces
cerevisiae. In one embodiment the plasmid may or may not be a 2
.mu.m-family plasmid and the host cell will be compatible with the
2 .mu.m-family plasmid used (see below for a full description of
the following plasmids). For example, where the plasmid is based on
pSR1, pSB3 or pSB4 then a suitable yeast cell is Zygosaccharomyces
rouxii; where the plasmid is based on pSB1 or pSB2 then a suitable
yeast cell is Zygosaccharomyces bailli; where the plasmid is based
on pSM1 then a suitable yeast cell is Zygosaccharomyces fermentati;
where the plasmid is based on pKD1 then a suitable yeast cell is
Kluyveromyces drosophilarum; where the plasmid is based on pPM1
then a suitable yeast cell is Pichia membranaefaciens; where the
plasmid is based on the 2 .mu.m plasmid then a suitable yeast cell
is Saccharomyces cerevisiae or Saccharomyces carlsbergensis. Thus,
the plasmid may be based on the 2 .mu.m plasmid and the yeast cell
may be Saccharomyces cerevisiae. A 2 .mu.m-family plasmid can be
said to be "based on" a naturally occurring plasmid if it comprises
one, two or preferably three of the genes FLP, REP1 and REP2 having
sequences derived from that naturally occurring plasmid.
[0332] A plasmid as defined above, may be introduced into a host
through standard techniques. With regard to transformation of
prokaryotic host cells, see, for example, Cohen et al (1972) Proc.
Natl. Acad. Sci. USA 69, 2110 and Sambrook et al (2001) Molecular
Cloning, A Laboratory Manual, 3.sup.rd Ed. Cold Spring Harbor
Laboratory, Cold Spring Harbor, N.Y. Transformation of yeast cells
is described in Sherman et al (1986) Methods In Yeast Genetics, A
Laboratory Manual, Cold Spring Harbor, N.Y. The method of Beggs
(1978) Nature 275, 104-109 is also useful. Methods for the
transformation of S. cerevisiae are taught generally in EP 251 744,
EP 258 067 and WO 90/01063, all of which are incorporated herein by
reference. With regard to vertebrate cells, reagents useful in
transfecting such cells, for example calcium phosphate and
DEAF-dextran or liposome formulations, are available from
Stratagene Cloning Systems, or Life Technologies Inc.,
Gaithersburg, Md. 20877, USA.
[0333] Electroporation is also useful for transforming cells and is
well known in the art for transforming fungal (including yeast)
cell, plant cells, bacterial cells and animal (including
vertebrate) cells. Methods for transformation of yeast by
electroporation are disclosed in Becker & Guarente (1990)
Methods Enzymol. 194, 182.
[0334] Generally, the plasmid will transform not all of the hosts
and it will therefore be necessary to select for transformed host
cells. Thus, a plasmid may comprise a selectable marker, including
but not limited to bacterial selectable marker and/or a yeast
selectable marker. A typical bacterial selectable marker is the
.beta.-lactamase gene although many others are known in the art.
Typical yeast selectable marker include LEU2, TRP1, HIS3, HIS4,
URA3, URA5, SFA1, ADE2, MET15, LYS5, LYS2, ILV2, FBA1, PSE1, PDI1
and PGK1. Those skilled in the art will appreciate that any gene
whose chromosomal deletion or inactivation results in an unviable
host, so called essential genes, can be used as a selective marker
if a functional gene is provided on the plasmid, as demonstrated
for PGK1 in a pgk1 yeast strain (Piper and Curran, 1990, Curr.
Genet. 17, 119). Suitable essential genes can be found within the
Stanford Genome Database (SGD), (http:://db.yeastgenome.org). Any
essential gene product (e.g. PDI1, PSE1, PGK1 or FBA1) which, when
deleted or inactivated, does not result in an auxotrophic
(biosynthetic) requirement, can be used as a selectable marker on a
plasmid in a host cell that, in the absence of the plasmid, is
unable to produce that gene product, to achieve increased plasmid
stability without the disadvantage of requiring the cell to be
cultured under specific selective conditions. By "auxotrophic
(biosynthetic) requirement" we include a deficiency which can be
complemented by additions or modifications to the growth medium.
Therefore, preferred "essential marker genes" in the context of the
present application are those that, when deleted or inactivated in
a host cell, result in a deficiency which cannot be complemented by
additions or modifications to the growth medium. Additionally, a
plasmid may comprise more than one selectable marker.
[0335] One selection technique involves incorporating into the
expression vector a DNA sequence marker, with any necessary control
elements, that codes for a selectable trait in the transformed
cell. These markers include dihydrofolate reductase, G418, neomycin
or zeocin resistance for eukaryotic cell culture, and tetracycline,
kanamycin, ampicillin (i.e. .beta.-lactamase) or zeocin resistance
genes for culturing in E. coli and other bacteria. Zeocin
resistance vectors are available from Invitrogen. Alternatively,
the gene for such selectable trait can be on another vector, which
is used to co-transform the desired host cell.
[0336] Another method of identifying successfully transformed cells
involves growing the cells resulting from the introduction of a
plasmid, optionally to allow the expression of a recombinant
polypeptide (i.e. a polypeptide which is encoded by a
polynucleotide sequence on the plasmid and is heterologous to the
host cell, in the sense that that polypeptide is not naturally
produced by the host). Cells can be harvested and lysed and their
DNA or RNA content examined for the presence of the recombinant
sequence using a method such as that described by Southern (1975)
J. Mol. Biol. 98, 503 or Berent et al (1985) Biotech. 3, 208 or
other methods of DNA and RNA analysis common in the art.
Alternatively, the presence of a polypeptide in the supernatant of
a culture of a transformed cell can be detected using
antibodies.
[0337] In addition to directly assaying for the presence of
recombinant DNA, successful transformation can be confirmed by well
known immunological methods when the recombinant DNA is capable of
directing the expression of the protein. For example, cells
successfully transformed with an expression vector produce proteins
displaying appropriate antigenicity. Samples of cells suspected of
being transformed are harvested and assayed for the protein using
suitable antibodies.
[0338] Thus, in addition to the transformed host cells themselves,
the present invention also contemplates a culture of those cells,
preferably a monoclonal (clonally homogeneous) culture, or a
culture derived from a monoclonal culture, in a nutrient medium.
Alternatively, transformed cells may represent an
industrially/commercially or pharmaceutically useful product and
can be used without further purification or can be purified from a
culture medium and optionally formulated with a carrier or diluent
in a manner appropriate to their intended industrial/commercial or
pharmaceutical use, and optionally packaged and presented in a
manner suitable for that use. For example, whole cells could be
immobilised; or used to spray a cell culture directly on to/into a
process, crop or other desired target. Similarly; whole cell, such
as yeast cells can be used as capsules for a huge variety of
applications, such as fragrances, flavours and pharmaceuticals.
[0339] Transformed host cells may be cultured for a sufficient time
and under appropriate conditions known to those skilled in the art,
and in view of the teachings disclosed herein, to permit the
expression of the helper protein(s) and the protein product of
choice.
[0340] The culture medium may be non-selective or place a selective
pressure on the maintenance of a plasmid.
[0341] The thus produced protein product of choice may be present
intracellularly or, if secreted, in the culture medium and/or
periplasmic space of the host cell.
[0342] Accordingly, the present invention, also provides a method
for producing a protein product of choice, the method
comprising:
(a) providing a host cell of the invention comprising a
polynucleotide encoding protein product of choice as defined above;
and (b) growing the host cell (for example, culturing the host cell
in a culture medium); thereby to produce a cell culture or
recombinant organism comprising an increased level of the protein
product of choice compared to the level of production of the
protein product of choice achieved by growing (for example,
culturing), under the same conditions, the same host cell that has
not been genetically modified to cause over-expression of one or
more helper proteins.
[0343] The step of growing the host cell may or may not involve
allowing a host cell derived from a multicellular organism to be
regrown into a multicellular recombinant organism (such as a plant
or animal) and, optionally, producing one or more generations of
progeny therefrom.
[0344] The method may or may not further comprise the step of
purifying the thus expressed protein product of choice from the
cultured host cell, recombinant organism or culture medium.
[0345] The step of "purifying the thus expressed protein product of
choice from the cultured host cell, recombinant organism or culture
medium" optionally comprises cell immobilisation, cell separation
and/or cell breakage, but always comprises at least one other
purification step different from the step or steps of cell
immobilisation, separation and/or breakage.
[0346] Cell immobilisation techniques, such as encasing the cells
using calcium alginate bead, are well known in the art. Similarly,
cell separation techniques, such as centrifugation,
filtration.(e.g. cross-flow filtration, expanded bed chromatography
and the like) are well known in the art. Likewise, methods of cell
breakage, including beadmilling, sonication, enzymatic exposure and
the like are well known in the art.
[0347] The "at least one other purification step" may be any other
step suitable for protein purification known in the art. For
example purification techniques for the recovery of recombinantly
expressed albumin have been disclosed in: WO 92/04367, removal of
matrix-derived dye; EP 464 590, removal of yeast-derived colorants;
EP 319 067, alkaline precipitation and subsequent application of
the albumin to a lipophilic phase; and WO 96/37515, U.S. Pat. No.
5,728,553 and WO 00/44772, which describe complete purification
processes; all of which are incorporated herein by reference.
[0348] Proteins other than albumin may be purified from the culture
medium by any technique that has been found to be useful for
purifying such proteins.
[0349] Suitable methods include ammonium sulphate or ethanol
precipitation, acid or solvent extraction, anion or cation exchange
chromatography, phosphocellulose chromatography, hydrophobic
interaction chromatography, affinity chromatography, hydroxyapatite
chromatography, lectin chromatography, concentration, dilution, pH
adjustment, diafiltration, ultrafiltration, high performance liquid
chromatography ("HPLC"), reverse phase HPLC, conductivity
adjustment and the like.
[0350] In one embodiment, any one or more of the above mentioned
techniques may or may not be used to further purifying the thus
isolated protein to a commercially or industrially acceptable level
of purity. By commercially or industrially acceptable level of
purity, we include the provision of the protein at a concentration
of at least 10.sup.4 gL.sup.-1, 10.sup.-3 gL.sup.-1, 0.01
gL.sup.-1, 0.02 gL.sup.-1, 0.03 gL.sup.-1, 0.04 gL.sup.-1, 0.05
gL.sup.-1, 0.06 gL.sup.-1, 0.07 gL.sup.-1, 0.08 gL.sup.-1, 0.09
gL.sup.-1, 0.1 gL.sup.-1, 0.2 gL.sup.-1, 0.3 gL.sup.-1, 0.4
gL.sup.-1, 0.5 gL.sup.-1, 0.6 gL.sup.-1, 0.7 gL.sup.-1, 0.8
gL.sup.-1, 0.9 gL.sup.-1, 1 gL.sup.-1, 2 gL.sup.-1, 3 gL.sup.-1, 4
gL.sup.-1, 5 gL.sup.-1, 6 gL.sup.-1, 7 gL.sup.-1, 8 gL.sup.-1, 9
gL.sup.-1, 10 gL.sup.-1, 15 gL.sup.-1, 20 gL.sup.-1, 25 gL.sup.-1,
30 gL.sup.-1, 40 gL.sup.-1, 50 gL.sup.-1, 60 gL.sup.-1, 70
gL.sup.-1, 70 gL.sup.-1, 90 gL.sup.-1, 100 gL.sup.-1, 150
gL.sup.-1, 200 gL.sup.-1, 250 gL.sup.-1, 300 gL.sup.-1, 350
gL.sup.-1, 400 gL.sup.-1, 500 gL.sup.-1, 600 gL.sup.-1, 700
gL.sup.-1, 800 gL.sup.-1, 900 gL.sup.-1, 1000 gL.sup.-1, or
more.
[0351] A commercially or industrially acceptable level of purity
may be obtained by a relatively crude purification method by which
the protein product of choice is put into a form suitable for its
intended purpose. A protein preparation that has been purified to a
commercially or industrially acceptable level of purity may, in
addition to the protein product of choice, also comprise, for
example, cell culture components such as host cells or debris
derived therefrom. Alternatively, high molecular weight components
(such as host cells or debris derived therefrom) may or may not be
removed (such as by filtration or centrifugation) to obtain a
composition comprising the protein product of choice and,
optionally, a functionally acceptable level of low molecular weight
contaminants derived from the cell culture process.
[0352] The protein may or may not be purified to achieve a
pharmaceutically acceptable level of purity. A protein has a
pharmaceutically acceptable level of purity if it is essentially
pyrogen free and can be administered in a pharmaceutically
efficacious amount without causing medical effects not associated
with the activity of the protein.
[0353] The resulting protein may be used for any of its known
utilities, which, in the case of albumin, include i.v.
administration to patients to treat severe burns, shock and blood
loss, supplementing culture media, and as an excipient in
formulations of other proteins.
[0354] A method of the present invention may or may not further
comprise the step of formulating the purified protein product of
choice with a carrier or diluent and optionally presenting the thus
formulated protein in a unit dosage form.
[0355] Although it is possible for a therapeutically useful protein
obtained by a process of the invention to be administered alone, it
is preferable to present it as a pharmaceutical formulation,
together with one or more acceptable carriers or diluents. The
carrier(s) or diluent(s) must be "acceptable" in the sense of being
compatible with the desired protein and not deleterious to the
recipients thereof. Typically, the carriers or diluents will be
water or saline which will be sterile and pyrogen free.
[0356] Optionally the thus formulated protein will be presented in
a unit dosage form, such as in the form of a tablet, capsule,
injectable solution or the like.
[0357] Alternatively, a method of the present invention may or may
not further comprise the step of lyophilising the thus purified
protein product of choice.
Detailed Description of Helper Proteins
[0358] JEM1 is one S. cerevisiae helper protein of interest for the
present invention. It is also known as KAR8, and its gene is a
non-essential gene located on chromosome X. It is a DnaJ-like
chaperone and is thought to be required for nuclear membrane fusion
during mating. It localises to the ER membrane and exhibits genetic
interactions with Kar2p (described further below). A published
protein sequence for the protein Jem1p is as follows:
TABLE-US-00001
MILISGYCLLVYSVILPVLISASKLCDLAELQRLNKNLKVDTESLPKYQWIAGQLEQNCM
TADPASENMSDVIQLANQIYYKIGLIQLSNDQHLRAINTFEKIVFNETYKGSFGKLAEKR
LQELYVDFGMWDKVHQKDDQYAKYLSLNETIRNKISSKDVSVEEDISELLRITPYDVNVL
STHIDVLFHKLAEEIDVSLAAAIILDYETILDKHLASLSIDTRLSIHYVISVLQTFVLNS
DASFNIRKCLSIDMDYDKCKKLSLTISKLNKVNPSKRQILDPATYAFENKKFRSWDRIIE
FYLKDKKPFITPMKILNKDTNFKNNYFFLEEIIKQLIEDVQLSRPLAKNLFEDPPITDGF
VKPKSYYHTDYLVYIDSILCQASSMSPDVKRAKLAAPFCKKSLRHSLTLETWKHYQDAKS
EQKPLPETVLSDVWNSNPHLLMYMVNSILNKSRSKPHSQFKKQLYDQINKFFQDNGLSES
TNPYVMKNFRLLQKQLQTYKEHKHRNFNQQYFQQQQQQQQHQRHQAPPAAPNYDPKKDYY
KILGVSPSASSKEIRKAYLNLTKKYHPDKIKANHNDKQESIHETMSQINEAYETLSDDDK
RKEYDLSRSNPRRNTFPQGPRQNNMFKNPGSGFPFGNGFKMNFGL*
[0359] The ORF of the JEM1 gene is 1.938 kbp in size. A published
nucleotide coding sequence of JEM1 is as follows, although it will
be appreciated that the sequence on be modified by degenerate
substitutions to obtain alternative nucleotide sequences which
encode an identical protein product:
TABLE-US-00002
ATGATACTGATCTCGGGATACTGTCTTTTAGTGTATAGCGTTATTTTGCCAGTACTGATA
TCGGCTTCTAAGTTATGTGATTTGGCTGAGTTACAACGATTGAACAAGAATTTAAAAGTA
GACACTGAATCCTTGCCAAAATACCAATGGATCGCTGGGCAGTTGGAACAAAACTGCATG
ACTGCGGATCCAGCAAGTGAAAATATGTCAGACGTAATTCAACTAGCCAATCAAATATAC
TACAAAATTGGGCTGATCCAATTATCCAACGATCAACATCTAAGAGCTATTAACACATTT
GAAAAAATCGTTTTTAATGAAACTTACAAAGGTTCTTTTGGGAAGCTGGCGGAAAAGAGG
CTACAAGAGCTGTATGTCGATTTTGGGATGTGGGACAAGGTGCATCAGAAGGATGATCAG
TATGCGAAATATCTGTCCTTGAATGAAACCATCAGAAACAAAATATCATCCAAAGACGTT
TCTGTGGAGGAAGATATTTCTGAGCTGCTACGCATAACGCCGTACGATGTTAACGTCCTC
TCCACGCACATCGATGTTCTTTTTCACAAACTAGCTGAAGAAATTGACGTTTCGTTAGCT
GCTGCTATCATTTTGGATTACGAAACAATCCTCGACAAGCATTTGGCTAGCTTAAGCATA
GATACAAGACTTTCGATTCATTATGTCATATCTGTTTTACAGACCTTTGTACTTAACTCA
GATGCGTCGTTCAATATAAGAAAATGCCTTTCCATTGATATGGACTATGATAAATGTAAA
AAACTAAGCCTGACTATTTCCAAATTGAACAAGGTGAATCCATCAAAAAGACAGATCCTG
GATCCAGCAACATATGCATTTGAGAACAAAAAGTTTAGAAGTTGGGATAGAATTATTGAA
TTTTATTTGAAGGATAAGAAGCCATTTATTACACCAATGAAAATTCTTAACAAAGATACA
AACTTTAAAAACAACTACTTCTTTTTAGAGGAAATTATCAAACAATTGATAGAAGACGTT
CAACTGTCGAGACCTTTGGCAAAAAATTTATTCGAAGATCCCCCAATAACCGATGGTTTT
GTCAAACCAAAATCATACTATCATACCGATTATCTAGTATACATTGATTCCATTCTTTGT
CAGGCTTCTAGCATGAGTCCGGACGTCAAGAGAGCTAAACTGGCTGCGCCGTTCTGTAAA
AAGAGTTTGAGGCATTCACTAACACTAGAAACATGGAAACACTATCAGGATGCTAAGTCC
GAGCAAAAACCTTTACCTGAGACGGTATTGAGTGATGTATGGAATTCCAATCCTCATTTG
CTGATGTATATGGTAAACTCAATACTTAATAAAAGTAGGTCTAAACCTCATTCACAGTTC
AAAAAGCAATTATATGACCAGATAAACAAATTTTTCCAAGATAACGGCCTCTCAGAGTCG
ACCAATCCATACGTGATGAAGAACTTCCGATTATTACAGAAACAATTACAAACCTATAAA
GAGCATAAACATCGGAATTTCAACCAGCAATATTTCCAACAACAACAACAGCAGCAACAA
CACCAACGACATCAAGCACCCCCAGCAGCGCCTAACTACGACCCAAAAAAGGACTATTAT
AAAATTCTTGGAGTATCGCCTAGTGCTAGTTCGAAAGAAATAAGGAAAGCATATTTAAAT
TTAACCAAAAAATACCACCCAGACAAAATAAAGGCCAACCATAACGACAAACAAGAATCA
ATTCACGAAACTATGTCACAAATCAATGAAGCGTACGAAACATTAAGTGATGACGATAAA
AGGAAGGAATACGATCTTTCCAGATCAAACCCCCGCCGCAACACTTTTCCTCAGGGGCCT
AGGCAAAATAACATGTTCAAAAATCCAGGAAGTGGCTTCCCATTCGGAAATGGCTTTAAA
ATGAATTTTGGGCTTTGA
[0360] Further information concerning JEM1 can be seen at the
following URL address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000003609.
[0361] It will be appreciated that, by "JEM1", we include fragments
or variants thereof having equivalent JEM1-like activity. Such
variants may or may not include bacterial DnaJ proteins and/or may
or may not include eukaryotic DnaJ type proteins, such as other
members of the Hsp40 family. In one embodiment, a variant of JEM1
may not be SCJ1.
[0362] LHS1 is another S. cerevisiae helper protein of interest for
the present invention. It is also known as CER1 or SSI1, is encoded
by a non-essential gene which is located on chromosome XI. It is
thought to be a molecular chaperone of the endoplasmic reticulum
lumen, involved in polypeptide translocation and folding. It is a
member of the HSP70 family, localizes to the lumen of the ER, and
is thought to be regulated by the unfolded protein response
pathway.
[0363] A published protein sequence for the protein Lhs1p is as
follows:
TABLE-US-00003
MRNVLRLLFLTAFVAIGSLAAVLGVDYGQQNIKAIVVSPQAPLELVLTPEAKRKEISGLS
IKRLPGYGKDDPNGIERIYGSAVGSLATRFPQNTLLHLKPLLGKSLEDETTVTLYSKQHP
GLEMVSTNRSTIAFLVDNVEYPLEELVAMNVQEIANRANSLLKDRDARTEDFVNKMSFTI
PDFFDQHQRKALLDASSITTGIEETYLVSEGMSVAVNFVLKQRQFPPGEQQHYIVYDMGS
GSIKASMFSILQPEDTTQPVTIEFEGYGYNPHLGGAKFTMDIGSLIENKFLETHPAIRTD
ELHANPKALAKINQAAEKAKLILSANSEASINIESLINDIDFRTSITRQEFEEFIADSLL
DIVKPINDAVTKQFGGYGTNLPEINGVILAGGSSRIPIVQDQLIKLVSEEKVLRNVNADE
SAVNGVVMRGIKLSNSFKTKPLNVVDRSVNTYSFKLSNESELYDVFTRGSAYPNKTSILT
NTTDSIPNNFTIDLFENGKLFETITVNSGAIKNSYSSDKCSSGVAYNITFDLSSDRLFSI
QEVNCICQSENDIGNSKQIKNKGSRLAFTSEDVEIKRLSPSERSRLHEHIKLLDKQDKER
FQFQENLNVLESNLYDARNLLMDDEVMQNGPKSQVEELSEMVKVYLDWLEDASFDTDPED
IVSRIREIGILKKKIELYMDSAKEPLNSQQFKGMLEEGHKLLQAIETHKNTVEEFLSQFE
TEFADTIDNVREEFKKIKQPAYVSKALSTWEETLTSFKNSISEIEKFLAKNLFGEDLREH
LFEIKLQFDMYRTKLEEKLRLIKSGDESRLNEIKKLHLRNFRLQKRKEEKLKRKLEQEKS
RNNNETESTVINSADDKTTIVNDKTTESNPSSEEDILHDEL*
[0364] The ORF of the LHS1 gene is 2.646 kbp in size. A published
nucleotide coding sequence of LHS1 is as follows, although it will
be appreciated that the sequence can be modified by degenerate
substitutions to obtain alternative nucleotide sequences which
encode an identical protein product:
TABLE-US-00004
ATGCGAAACGTTTTAAGGCTTTTATTTTTAACAGCTTTTGTTGCTATAGGGTCTTTAGCA
GCCGTTTTAGGTGTTGATTACGGTCAGCAAAATATCAAGGCCATTGTGGTTTCTCCGCAA
GCCCCATTAGAACTTGTGCTCACACCAGAGGCAAAACGGAAGGAGATATCTGGTCTTTCG
ATAAAAAGATTACCAGGTTATGGAAAGGATGATCCGAATGGGATTGAAAGAATCTACGGT
TCCGCTGTTGGCAGTTTAGCAACAAGGTTTCCCCAAAACACATTGTTGCATTTGAAACCG
CTACTTGGGAAATCACTAGAAGATGAAACCACTGTAACTTTGTATTCAAAACAACACCCC
GGTTTAGAAATGGTATCAACAAATAGAAGTACCATAGCCTTTTTAGTTGATAATGTGGAA
TATCCATTGGAAGAGTTAGTGGCAATGAATGTCCAAGAGATTGCCAATAGAGCCAATTCA
CTGTTGAAGGATAGAGATGCAAGAACTGAGGACTTTGTAAACAAGATGAGTTTTACAATT
CCTGACTTTTTTGACCAACATCAAAGGAAAGCACTTTTAGATGCCAGTTCAATAACCACA
GGAATCGAAGAGACATATCTGGTTAGTGAAGGGATGTCTGTTGCAGTTAACTTTGTATTA
AAGCAGCGCCAATTTCCACCAGGTGAACAGCAGCATTATATCGTATATGACATGGGGAGC
GGTTCTATTAAGGCCTCAATGTTCTCTATATTGCAGCCGGAGGACACTACTCAGCCCGTT
ACAATAGAATTTGAAGGATATGGGTATAATCCACATCTAGGTGGTGCAAAGTTTACAATG
GATATTGGCAGTTTGATAGAGAATAAGTTTTTGGAAACACACCCAGCCATAAGAACTGAT
GAATTGCACGCTAATCCCAAGGCCTTAGCAAAAATCAACCAAGCAGCAGAGAAGGCAAAG
TTAATTTTAAGCGCCAATTCTGAGGCAAGTATTAACATAGAATCACTGATCAACGATATT
GATTTCCGTACTTCTATAACTAGACAGGAATTCGAAGAATTTATTGCAGACTCGTTATTG
GACATTGTCAAACCCATAAATGACGCTGTTACAAAACAATTCGGTGGCTATGGAACAAAT
TTACCTGAGATAAATGGGGTCATTTTGGCGGGAGGCTCTTCCCGAATTCCCATTGTGCAG
GATCAATTAATCAAACTCGTATCCGAAGAAAAAGTGTTGAGAAATGTCAATGCTGATGAA
TCAGCTGTGAATGGTGTTGTTATGAGAGGGATCAAGTTATCTAATTCGTTTAAGACCAAG
CCGTTAAATGTTGTTGACCGTTCTGTAAATACTTATTCATTCAAATTATCAAACGAATCT
GAACTGTATGATGTGTTCACGCGCGGAAGTGCTTATCCAAACAAAACATCTATTTTGACA
AACACGACTGATTCGATTCCTAATAATTTTACCATTGACTTATTTGAGAATGGTAAATTG
TTCGAAACTATCACAGTTAATTCAGGAGCTATAAAGAATTCATATTCCTCTGATAAGTGC
TCGTCAGGAGTTGCGTATAACATTACTTTCGACTTGTCCAGTGATAGATTATTCTCTATT
CAAGAGGTTAACTGCATTTGTCAGAGCGAAAATGACATAGGTAACTCCAAGCAAATTAAG
AACAAAGGCAGCCGTTTGGCTTTTACTTCTGAGGATGTTGAGATCAAAAGGCTTTCTCCT
TCAGAACGTTCGCGTTTGCATGAGCATATCAAGTTGCTCGATAAACAGGATAAGGAAAGA
TTTTCAATTCCAAGAAAATTTAAACGTTCTTGAAAGTAACTTGTATGATGCTAGAAACCTG
CTAATGGATGATGAAGTTATGCAAAATGGACCAAAATCCCAAGTAGAAGAGTTATCGGAG
ATGGTTAAAGTATATTTGGATTGGCTCGAAGATGCATCCTTTGATACTGACCCTGAGGAT
ATAGTTAGCAGAATTAGAGAAATTGGAATATTAAAAAAGAAAATAGAACTTTACATGGAT
TCTGCAAAGGAACCTTTGAACTCTCAACAATTTAAAGGAATGCTTGAAGAAGGCCATAAG
TTACTTCAGGCTATAGAAACCCATAAGAATACCGTTGAAGAATTTTTGAGTCAATTTGAA
ACCGAGTTTGCGGATACCATAGATAATGTTAGAGAAGAATTTAAAAAGATTAAGCAACCA
GCGTATGTGTCGAAGGCGTTATCTACATGGGAGGAAACCTTAACCTCTTTTAAAAATTCC
ATTAGCGAAATAGAGAAGTTCCTGGCAAAAAACCTATTTGGCGAAGACCTTCGTGAACAT
TTATTTGAAATCAAATTACAATTTGATATGTATCGTACGAAACTAGAGGAAAAACTGCGT
TTAATAAAAAGCGGTGATGAAAGTCGCTTAAATGAAATAAAGAAGTTACATTTAAGAAAC
TTCCGCCTACAAAAGAGAAAGGAGGAAAAGTTGAAAAGAAAGCTTGAACAGGAAAAAAGC
AGAAACAACAATGAAACAGAATCGACAGTAATCAACTCGGCTGACGATAAAACTACTATT
GTCAATGACAAGACCACCGAGTCGAATCCAAGTTCTGAGGAAGACATTTTGCATGATGAA
TTATAG
[0365] Further information on LHS1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000001556.
[0366] It will be appreciated that, by "LHS1", we include fragments
or variants thereof having equivalent LHS1-like activity. Such
variants may or may not include bacterial DnaK proteins and/or
eukaryotic DnaK type proteins, such as other members of the Hsp70
family.
[0367] SCJ1 is another S. cerevisiae helper protein of interest for
the present invention. It is one of several homologs of bacterial
chaperone DnaJ, located in the ER lumen where it cooperates with
Kar2p (described below) to mediate maturation of proteins.
[0368] A published protein sequence for the protein Scj1p is as
follows:
TABLE-US-00005 MIPKLYIHLILSLLLLPLILAQDYYAILEIDKDATEKEIKSAYRQLSKKY
HPDKNAGSEEAHQKFIEVGEAYDVLSDPEKKKIYDQFGADAVKNGGGGGG
PGGPGAGGFHDPFDIFERMFQGGHGGPGGGFGQRQRQRGPMIKVQEKLSL
KQFYSGSSIEFTLNLNDECDACHGSGSADGKLAQCPDCQGRGVIIQVLRM
GIMTQQIQQMCGRCGGTGQIIKNECKTCHGKKVTKKNKFFHVDVPPGAPR
NYMDTRVGEAEKGPDFDAGDLVIEFKEKDTENMGYRRRGDNLYRTEVLSA
AEALYGGWQRTIEFLDENKPVKLSRPAHVVVSNGEVEVVKGFGMPKGSKG
YGDLYIDYVVVMPKTFKSGQNMLKDEL*
[0369] SCJ1 is encoded by a non-essential gene comprising an ORF of
1.134 kbp. The gene is located on chromosome XIII. A published
nucleotide coding sequence of SCJ1 is as follows, although it will
be appreciated that the sequence can be modified by degenerate
substitutions to obtain alternative nucleotide sequences which
encode an identical protein product:
TABLE-US-00006
ATGATTCCAAAATTATATATACATTTGATACTATCTTTATTGTTGTTGCCGCTAATTTTG
GCGCAGGATTATTATGCAATACTAGAGATAGACAAAGATGCCACTGAGAAGGAAATCAAA
TCAGCGTACAGACAATTGTCTAAGAAGTACCATCCGGATAAAAATGCTGGGAGCGAAGAA
GCCCATCAAAAATTCATTGAAGTCGGCGAGGCATACGATGTATTGAGCGATCCTGAAAAG
AAAAAGATTTATGACCAGTTTGGTGCAGATGCTGTAAAGAATGGCGGTGGCGGTGGCGGT
CCAGGAGGCCCTGGCGCAGGTGGATTCCACGATCCGTTTGACATATTCGAACGGATGTTT
CAAGGAGGTCATGGAGGTCCTGGCGGCGGATTTGGCCAGAGACAGAGGCAGCGTGGTCCA
ATGATCAAGGTCCAGGAAAAACTATCTTTAAAGCAGTTTTATTCCGGGTCCTCGATAGAA
TTTACTTTAAACCTAAACGATGAATGTGATGCATGCCATGGTAGTGGCTCTGCAGATGGT
AAGCTGGCCCAATGTCCCGATTGTCAAGGTCGTGGGGTTATAATACAAGTGCTGCGCATG
GGTATTATGACGCAGCAGATTCAACAGATGTGTGGTAGGTGTGGTGGTACGGGACAAATT
ATCAAAAATGAATGCAAAACATGTCACGGCAAAAAAGTTACCAAAAAGAACAAGTTCTTC
CACGTTGACGTTCCACCAGGCGCACCAAGAAACTACATGGACACAAGAGTCGGCGAGGCT
GAAAAAGGGCCTGACTTTGACGCCGGTGACTTGGTCATAGAATTCAAGGAAAAGGATACT
GAGAACATGGGTTACAGAAGAAGAGGCGACAATCTGTACAGAACAGAAGTTCTTTCTGCT
GCGGAAGCGCTATACGGCGGATGGCAAAGAACGATAGAATTCCTTGATGAGAACAAGCCC
GTTAAGTTATCTAGACCCGCTCATGTAGTTGTCTCCAATGGCGAAGTTGAAGTCGTGAAG
GGATTCGGCATGCCCAAGGGTAGCAAGGGTTACGGTGATTTGTACATAGACTACGTCGTT
GTCATGCCAAAGACTTTCAAATCTGGGCAAAATATGCTCAAAGATGAGTTGTAG
[0370] Further information on SCJ1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S006004827.
[0371] It will be appreciated that, by "SCJ1", we include fragments
or variants thereof having equivalent SCJ1-like activity.
[0372] KAR2 is another S. cerevisiae helper protein of interest for
the present invention. KAR2 is also known as BIP or GRP78. Kar2p,
is an ATPase involved in protein import into the ER. Kar2p also
acts as a chaperone to mediate protein folding in the ER and may
play a role in ER export of soluble proteins. It is also thought to
regulate the unfolded protein response via interaction with kelp. A
published protein sequence for the protein Kar2p is as follows:
TABLE-US-00007 MFFNRLSAGKLLVPLSVVLYALFVVILPLQNSFHSSNVLVRGADDVENYG
TVIGIDLGTTYSCVAVMKNGKTEILANEQGNRITPSYVAFTDDERLIGDA
AKNQVAANPQNTIFDIKRLIGLKYNDRSVQKDIKHLPFNVVNKDGKPAVE
VSVKGEKKVFTPEEISGMILGKMKQIAEDYLGTKVTHAVVTVPAYFNDAQ
RQATKDAGTIAGLNVLRIVNEPTAAAIAYGLDKSDKEHQIIVYDLGGGTF
DVSLLSIENGVFEVQATSGDTHLGGEDFDYKIVRQLIKAFKKKHGIDVSD
NNKALAKLKREAEKAKRALSSQMSTRIEIDSFVDGIDLSETLTRAKFEEL
NLDLFKKTLKPVEKVLQDSGLEKKDVDDIVLVGGSTRIPKVQQLLESYFD
GKKASKGINPDEAVAYGAAVQAGVLSGEEGVEDIVLLDVNALTLGIETTG
GVMTPLIKRNTAIPTKKSQIFSTAVDNQPTVMIKVYEGERAMSKDNNLLG
KFELTGIPPAPRGVPQIEVTFALDANGILKVSATDKGTGKSESITITNDK
GRLTQEEIDRMVEEAEKFASEDASIKAKVESRNKLENYAHSLKNQVNGDL
GEKLEEEDKETLLDAANDVLEWLDDNFETAIAEDFDEKFESLSKVAYPIT
SKLYGGADGSGAADYDDEDEDDDGDYFEHDEL*
[0373] KAR2 is encoded by an essential gene comprising an ORF that
is 2.049 kbp in size and located on chromosome X. A published
nucleotide coding sequence of KAR2 is as follows, although it will
be appreciated that the sequence can be modified by degenerate
substitutions to obtain alternative nucleotide sequences which
encode an identical protein product:
TABLE-US-00008
ATGTTTTTCAACAGACTAAGCGCTGGCAAGCTGCTGGTACCACTCTCCGTGGTCCTGTAC
GCCCTTTTCGTGGTAATATTACCTTTACAGAATTCTTTCCACTCCTCCAATGTTTTAGTT
AGAGGTGCCGATGATGTAGAAAACTACGGAACTGTTATCGGTATTGACTTAGGTACTACT
TATTCCTGTGTTGCTGTGATGAAAAATGGTAAGACTGAAATTCTTGCTAATGAGCAAGGT
AACAGAATCACCCCATCTTACGTGGCATTCACCGATGATGAAAGATTGATTGGTGATGCT
GCAAAGAACCAAGTTGCTGCCAATCCTCAAAACACCATCTTCGACATTAAGAGATTGATC
GGTTTGAAATATAACGACAGATCTGTTCAGAAGGATATCAAGCACTTGCCATTTAATGTG
GTTAATAAAGATGGGAAGCCCGCTGTAGAAGTAAGTGTCAAAGGAGAAAAGAAGGTTTTT
ACTCCAGAAGAAATTTCTGGTATGATCTTGGGTAAGATGAAACAAATTGCCGAAGATTAT
TTAGGCACTAAGGTTACCCATGCTGTCGTTACTGTTCCTGCTTATTTCAATGACGCGCAA
AGACAAGCCACCAAGGATGCTGGTACCATCGCTGGTTTGAACGTTTTGAGAATTGTTAAT
GAACCAACCGCAGCCGCCATTGCCTACGGTTTGGATAAATCTGATAAGGAACATCAAATT
ATTGTTTATGATTTGGGTGGTGGTACTTTCGATGTCTCTCTATTGTCTATTGAAAACGGT
GTTTTCGAAGTCCAAGCCACTTCTGGTGATACTCATTTAGGTGGTGAAGATTTTGACTAT
AAGATCGTTCGTCAATTGATAAAAGCTTTCAAGAAGAAGCATGGTATTGATGTGTCTGAC
AACAACAAGGCCCTAGCTAAATTGAAGAGAGAAGCTGAAAAGGCTAAACGTGCCTTGTCC
AGCCAAATGTCCACCCGTATTGAAATTGACTCCTTCGTTGATGGTATCGACTTAAGTGAA
ACCTTGACCAGAGCTAAGTTTGAGGAATTAAACCTAGATCTATTCAAGAAGACCTTGAAG
CCTGTCGAGAAGGTTTTGCAAGATTCTGGTTTGGAAAAGAAGGATGTTGATGATATCGTT
TTGGTTGGTGGTTCTACTAGAATTCCAAAGGTCCAACAATTGTTAGAATCATACTTTGAT
GGTAAGAAGGCCTCCAAGGGTATTAACCCAGATGAAGCTGTTGCATACGGTGCAGCCGTT
CAAGCTGGTGTCTTATCCGGTGAAGAAGGTGTCGAAGATATTGTTTTATTGGATGTCAAC
GCTTTGACTCTTGGTATTGAAACCACTGGTGGTGTCATGACTCCATTAATTAAGAGAAAT
ACTGCTATTCCTACAAAGAAATCCCAAATTTTCTCTACTGCCGTTGACAACCAACCAACC
GTTATGATCAAGGTATACGAGGGTGAAAGAGCCATGTCTAAGGACAACAATCTATTAGGT
AAGTTTGAATTAACCGGCATTCCACCAGCACCAAGAGGTGTACCTCAAATTGAAGTCACA
TTTGCACTTGACGCTAATGGTATTCTGAAGGTGTCTGCCACAGATAAGGGAACTGGTAAA
TCCGAATCTATCACCATCACTAACGATAAAGGTAGATTAACCCAAGAAGAGATTGATAGA
ATGGTTGAAGAGGCTGAAAAATTCGCTTCTGAAGACGCTTCTATCAAGGCCAAGGTTGAA
TCTAGAAACAAATTAGAAAACTACGCTCACTCTTTGAAAAACCAAGTTAATGGTGACCTA
GGTGAAAAATTGGAAGAAGAAGACAAGGAAACCTTATTAGATGCTGCTAACGATGTTTTA
GAATGGTTAGATGATAACTTTGAAACCGCCATTGCTGAAGACTTTGATGAAAAGTTCGAA
TCTTTGTCCAAGGTCGCTTATCCAATTACTTCTAAGTTGTACGGAGGTGCTGATGGTTCT
GGTGCCGCTGATTATGACGACGAAGATGAAGATGACGATGGTGATTATTTCGAACACGAC
GAATTGTAG
[0374] Further information on KAR2 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000003571.
[0375] It will be appreciated that, by "KAR2", we include fragments
or variants thereof having equivalent KAR2-like activity.
[0376] SIL1 is another S. cerevisiae helper protein of interest for
the present invention and is also known as SLS1. In particular,
this helper protein was generally referred to as SLS1 in UK patent
application no. 0512707.1, from which this application claims
priority; it will be understood by the person skilled in the art
that reference in UK patent application no. 0512707.1 to SLS1 and
reference in this application to SIL1 should be taken to be
reference to the same helper protein. SIL1p is an ER-localized
protein required for protein translocation into the ER, which
interacts with the ATPase domain of the Kar2p chaperone suggesting
some role in modulating its activity. It is also thought to be a
homolog of Yarrowia lipolytica SIL1 and a GrpE-like protein in the
ER. A published protein sequence for the protein SIL1p is as
follows:
TABLE-US-00009 MVRILPIILSALSSKLVASTILHSSIHSVPSGGEIISAEDLKELEISGNS
ICVDNRCYPKIFEPRHDWQPILPGQELPGGLDIRINMDTGLKEAKLNDEK
NVGDNGSHELIVSSEDMKASPGDYEFSSDFKEMRNIIDSNPTLSSQDIAR
LEDSFDRIMEFAHDYKHGYKIITHEFALLANLSLNENLPLTLRELSTRVI
TSCLRNNPPVVEFINESFPNFKSKIMAALSNLNDSNHRSSNILIKRYLSI
LNELPVTSEDLPIYSTVVLQNVYERNNKDKQLQIKVLELISKILKADMYE
NDDTNLILFKRNAENWSSNLQEWANEFQEMVQNKSIDELHTRTFFDTLYN
LKKIEFSDITINKGFLNWLAQQCKARQSNLDNGLQERDTEQDSFDKKLID
SRHLIFGNPMAHRIKNFRDEL*
[0377] SIL1 is encoded by a non-essential gene comprising an ORF
that is 1.226 kbp in size and is located on chromosome XV. A
published nucleotide coding sequence of SIL1 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00010
ATGGTCCGGATTCTTCCCATAATTTTGAGCGCCCTATCTTCGAAATTAGTGGCGAGTACA
ATATTGCATTCATCCATACACTCAGTGCCATCTGGAGGCGAAATCATATCTGCAGAAGAT
CTTAAAGAACTTGAAATTTCAGGGAATTCGATCTGCGTTGATAATCGTTGCTATCCTAAG
ATATTTGAACCAAGACACGATTGGCAGCCCATACTGCCAGGTCAAGAACTCCCCGGTGGT
TTGGACATTAGAATAAACATGGACACAGGTTTAAAAGAGGCAAAACTAAATGATGAGAAG
AATGTCGGTGATAATGGTAGCCATGAGTTAATTGTATCTTCAGAAGACATGAAAGCATCG
CCTGGTGACTATGAATTTTCCAGTGATTTCAAAGAAATGAGAAACATCATAGATTCTAAC
CCGACTTTATCTTCACAGGACATTGCCAGATTGGAGGATAGTTTTGATAGAATAATGGAA
TTTGCGCATGATTACAAGCACGGCTACAAAATTATTACCCATGAATTCGCCCTCTTGGCC
AACCTTAGTCTCAATGAAAATTTGCCGTTAACATTGAGAGAGCTCAGTACTAGAGTCATT
ACCAGCTGCTTGAGAAACAATCCTCCTGTAGTCGAGTTCATTAATGAAAGTTTTCCAAAT
TTTAAAAGCAAAATCATGGCCGCTCTGTCAAATTTGAATGATTCTAACCACAGATCCTCT
AATATCCTAATAAAAAGATACTTGTCCATTTTAAACGAATTACCTGTCACATCCGAAGAT
CTTCCTATATACTCTACGGTTGTTTTACAAAATGTATATGAAAGAAACAACAAGGACAAA
CAGTTACAAATAAAAGTCCTGGAGTTGATCAGCAAAATTTTGAAGGCCGACATGTACGAA
AATGACGATACAAATCTAATTTTGTTCAAAAGAAATGCTGAGAATTGGTCGTCAAATCTG
CAAGAGTGGGCAAACGAGTTCCAAGAGATGGTCCAGAACAAAAGTATAGATGAACTACAT
ACAAGAACGTTTTTTGACACCCTTTACAACTTGAAGAAAATTTTCAAAAGTGACATCACG
ATCAACAAAGGGTTTTTGAATTGGTTAGCGCAACAATGTAAAGCCAGGCAATCTAACTTG
GACAATGGGCTCCAAGAGAGAGATACTGAACAAGACTCATTTGATAAGAAACTTATCGAC
AGCAGACACTTGATCTTTGGCAACCCCATGGCTCATAGAATAAAAAATTTCAGAGATGAA
CTCTGA
[0378] Further information on SIL1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000005391.
[0379] It will be appreciated that, by "SIL1", we include fragments
or variants (including homologues) thereof having equivalent
SIL1-like activity. In one embodiment, variants of SIL1 may or may
not include bacterial GrpE type proteins and/or animal (such as
mammalian) GrpE-like proteins. Variants of SIL1 may be a nucleotide
exchange factor for an Hsp70 family protein, which nucleotide
exchange factor is optionally not an Hsp70 family protein in
itself. Suitable variants of SIL1 may or may not be FES1 and/or
MGE1. A variant of SIL1 may or may not be localised to the lumen of
the ER (such as SIL1 itself) to the mitochondria (such as MGE1) or
to the cytosol (such as FES1). A variant of SIL1 may or may not
include proteins such as members so of the mammalian GrpE-like
protein family, the NEF family or BAG-1 (such as described in
Hohfeld and Jentsch (1997) EMBO J. 16, 6209), mammalian
BiP-associated protein (BAP) (Chung et al (2002) J. Biol. Chem.
277, 47557), a human GrpE-like protein (e.g. the protein defined by
accession number AAG31605) (Choglay et al (2001) Gene 267, 125), an
Arabidopsis thaliana GrpE-like protein (for example, accession
numbers AAK68792 and BAB08589) (Sato et al (1998) DNA Res. 5, 41),
a Chlamydia trachomatis Protein grpE (HSP-70 cofactor) (e.g.
accession number P36424), a Pongo pygmaeus adenine nucleotide
exchange factor (e.g. accession number CAH89792), a Mus musculus
mitochondrial GrpE-like 2 protein (e.g. accession number
NP.sub.--067271), a Mus musculus mitochondrial GrpE-like 1 protein
(e.g. accession number NP.sub.--077798), a Gallus gallus GrpE
protein homolog 2, mitochondrial precursor (Mt-GrpE#2) (e.g.
accession number XP.sub.--425191), a Gallus gallus BiP-associated
protein (e.g. accession number XP.sub.--414514), an Haemophilus
influenzae 86-028NP GrpE protein (e.g. as defined by accession
number YP.sub.--247735) (Harrison et al (2005) J. Bacteriol. 187,
4627), an Escherichia coli GrpE heat shock protein (e.g. as defined
by accession number NP.sub.--417104) (Riley et al (1997) Science
277, 1453), a Streptococcus pneumoniae GrpE heat shock protein
(e.g. as defined by accession number AAD23453), a Bacillus subtilis
GrpE protein accession number (e.g. as defined by BAA12463) (Mizuno
et al (1996) Microbiology (Reading, Engl.) 142, 3103) and/or a
Nicotiana tabacum chaperone GrpE type 1 or GrpE type 2 protein
(e.g. as defined by accession numbers AAC72386 or AAC72387)
(Padidam et al (1999) Plant Mol. Biol. 39, 871).
[0380] Variants of SIL1 may have an activity equivalent to SIL1,
when co-expressed with one or both of JEM1 and LHS1, for example in
the manner as set out in the present examples. Thus, a host cell of
the present invention, when genetically modified to cause
simultaneous over-expression of a variant of SILT with one or both
of JEM1 and LHS1, will provide at least substantially the same
increase in the production of a protein product and/or at least
substantially the same reduction of fragmentation of a protein
product, as is observed in the same host cell when genetically
modified to cause simultaneous over-expression of SIL1 with one or
both of JEM1 and LHS1, the increase being compared to the level of
production of the same protein product, and/or the level of
fragmentation of the same protein product, in the same host cell
that has not been genetically modified to cause overexpression of
any of LHS1, JEM1 or SIL1.
[0381] By "substantially the same increase in the production of a
protein product", we mean at least 10%, 20%, 30%, 40%, 50%, 60%,
70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, substantially 100% or
greater than 100% of the increase in production of a protein
product that is observed when the host cell is genetically modified
to cause simultaneous over-expression of SIL1 with one or both of
JEM1 and LHS1 (the increased being compared to the level of
production of the same protein product in the same host cell that
has not been genetically modified to cause overexpression of any of
LHS1, JEM1 or SIL1).
[0382] By "substantially the same reduction of fragmentation of a
protein product", we mean at least 10%, 20%, 30%, 40%, 50%, 60%,
70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, substantially 100% or
greater than 100% of the reduction of fragmentation of a protein
product that is observed when the host cell is genetically modified
to cause simultaneous over-expression of SIL1 with one or both of
JEM1 and LHS1 (the reduction of fragmentation of a protein product
being compared to the level of fragmentation of the same protein
product in the same host cell that has not been genetically
modified to cause overexpression of any of LHS1, JEM1 or SIL1).
[0383] FKB2 is another S. cerevisiae helper protein of interest for
the present invention and is also known as FPR2 and FKBP13. Fkb2p
is a membrane bound peptidyl-prolyl cis-trans isomerase (PPIase)
that binds to the drugs FK506 and rapamycin. The expression pattern
of Fkb2p suggests possible involvement in ER protein trafficking. A
published protein sequence for the protein Fkb2p is as follows:
TABLE-US-00011 MMFNIYLFVTFFSTILAGSLSDLEIGIIKRIPVEDCLIKAMPGDKVKVHY
TGSLLESGTVFDSSYSRGSPIAFELGVGRVIKGWDQGVAGMCVGEKRKLQ
IPSSLAYGERGVPGVIPPSADLVFDVELVDVKSAA*
[0384] FKB2 is encoded by a non-essential gene comprising an ORF
that is 0.408 kbp in size and is located on chromosome IV. A
published nucleotide coding sequence of FKB2 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00012 ATGATGTTTAATATTTACCTTTTCGTCACTTTTTTTTCCACCATTCTTGC
AGGTTCCCTGTCAGATTTGGAAATCGGTATTATCAAGAGAATACCGGTAG
AAGATTGCTTAATTAAGGCAATGCCAGGTGATAAAGTTAAGGTTCATTAT
ACAGGATCTTTATTAGAATCGGGAACTGTATTTGACTCAAGTTATTCAAG
AGGCTCTCCTATCGCTTTTGAACTTGGCGTTGGCAGAGTAATTAAAGGTT
GGGATCAAGGTGTTGCCGGCATGTGCGTTGGCGAAAAAAGAAAGCTGCAA
ATTCCAAGTTCTTTGGCCTACGGAGAAAGAGGTGTCCCAGGCGTCATTCC
TCCAAGTGCTGATTTGGTGTTTGATGTCGAATTGGTAGACGTGAAATCAG CCGCCTAG
[0385] Further information on FKB2 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000002927.
[0386] It will be appreciated that, by "FKB2", we include fragments
or variants thereof having equivalent FKB2-like activity.
[0387] SSA1 is another S. cerevisiae helper protein of interest for
the present invention and is also known as YG100. Ssa1p is an
ATPase that is involved in protein folding and nuclear localization
signal (NLS)-directed nuclear transport. It is a member of heat
shock protein 70 (HSP70) family. It forms a chaperone complex with
Ydj1p and is localized to the nucleus, cytoplasm, and cell wall A
published protein sequence for the protein Ssa1p is as follows:
TABLE-US-00013 MSKAVGIDLGTTYSCVAHFANDRVDIIANDQGNRTTPSFVAFTDTERLIG
DAAKNQAAMNPSNTVFDAKRLIGRNFNDPEVQADMKHFPFKLIDVDGKPQ
IQVEFKGETKNFTPEQISSMVLGKMKETAESYLGAKVNDAVVTVPAYFND
SQRQATKDAGTIAGLNVLRIINEPTAAAIAYGLDKKGKEEHVLIFDLGGG
TFDVSLLFIEDGIFEVKATAGDTHLGGEDFDNRLVNHFIQEFKRKNKKDL
STNQRALRRLRTACERAKRTLSSSAQTSVEIDSLFEGIDFYTSITRARFE
ELCADLERSTLDPVEKVLRDAKLDKSQVDEIVLVGGSTRIPKVQKLVTDY
FNGKEPNRSINPDEAVAYGAAVQAAILTGDESSKTQDLLLLDVAPLSLGI
ETAGGVMTKLIPRNSTISTKKFEIFSTYADNQPGVLIQVFEGERAKTKDN
NLLGKFELSGIPPAPRGVPQIEVTEDVDSNGILNVSAVEKGTGKSNKITI
TNDKGRLSKEDIEKMVAEAEKFKEEDEKESQRIASKNQLESIAYSLKNTI
SEAGDKLEQADKDTVTKKAEETISWLDSNTTASKEEFDDKLKELQDIANP
IMSKLYQAGGAPGGAAGGAPGGFPGGAPPAPEAEGPTVEEVD*
[0388] SSA1 is encoded by a non-essential gene comprising an ORF
that is 1.929 kbp in size and is located on chromosome I. A
published nucleotide coding sequence of SSA1 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00014
ATGTCAAAAGCTGTCGGTATTGATTTAGGTACAACATACTCGTGTGTTGCTCACTTTGCT
AATGATCGTGTGGACATTATTGCCAACGATCAAGGTAACAGAACCACTCCATCTTTTGTC
GCTTTCACTGACACTGAAAGATTGATTGGTGATGCTGCTAAGAATCAAGCTGCTATGAAT
CCTTCGAATACCGTTTTCGACGCTAAGCGTTTGATCGGTAGAAACTTCAACGACCCAGAA
GTGCAGGCTGACATGAAGCACTTCCCATTCAAGTTGATCGATGTTGACGGTAAGCCTCAA
ATTCAAGTTGAATTTAAGGGTGAAACCAAGAACTTTACCCCAGAACAAATCTCCTCCATG
GTCTTGGGTAAGATGAAGGAAACTGCCGAATCTTACTTGGGAGCCAAGGTCAATGACGCT
GTCGTCACTGTCCCAGCTTACTTCAACGATTCTCAAAGACAAGCTACCAAGGATGCTGGT
ACCATTGCTGGTTTGAATGTCTTGCGTATTATTAACGAACCTACCGCCGCTGCCATTGCT
TACGGTTTGGACAAGAAGGGTAAGGAAGAACACGTCTTGATTTTCGACTTGGGTGGTGGT
ACTTTCGATGTCTCTTTGTTGTTCATTGAAGACGGTATCTTTGAAGTTAAGGCCACCGCT
GGTGACACCCATTTGGGTGGTGAAGATTTTGACAACAGATTGGTCAACCACTTCATCCAA
GAATTCAAGAGAAAGAACAAGAAGGACTTGTCTACCAACCAAAGAGCTTTGAGAAGATTA
AGAACCGCTTGTGAAAGAGCCAAGAGAACTTTGTCTTCCTCCGCTCAAACTTCCGTTGAA
ATTGACTCTTTGTTCGAAGGTATCGATTTCTACACTTCCATCACCAGAGCCAGATTCGAA
GAATTGTGTGCTGACTTGTTCAGATCTACTTTGGACCCAGTTGAAAAGGTCTTGAGAGAT
GCTAAATTGGACAAATCTCAAGTCGATGAAATTGTCTTGGTCGGTGGTTCTACCAGAATT
CCAAAGGTCCAAAAATTGGTCACTGACTACTTCAACGGTAAGGAACCAAACAGATCTATC
AACCCAGATGAAGCTGTTGCTTACGGTGCTGCTGTTCAAGCTGCTATTTTGACTGGTGAC
GAATCTTCCAAGACTCAAGATCTATTGTTGTTGGATGTCGCTCCATTATCCTTGGGTATT
GAAACTGCTGGTGGTGTCATGACCAAGTTGATTCCAAGAAACTCTACCATTTCAACAAAG
AAGTTCGAGATCTTTTCCACTTATGCTGATAACCAACCAGGTGTCTTGATTCAAGTCTTT
GAAGGTGAAAGAGCCAAGACTAAGGACAACAACTTGTTGGGTAAGTTCGAATTGAGTGGT
ATTCCACCAGCTCCAAGAGGTGTCCCACAAATTGAAGTCACTTTCGATGTCGACTCTAAC
GGTATTTTGAATGTTTCCGCCGTCGAAAAGGGTACTGGTAAGTCTAACAAGATCACTATT
ACCAACGACAAGGGTAGATTGTCCAAGGAAGATATCGAAAAGATGGTTGCTGAAGCCGAA
AAATTCAAGGAAGAAGATGAAAAGGAATCTCAAAGAATTGCTTCCAAGAACCAATTGGAA
TCCATTGCTTACTCTTTGAAGAACACCATTTCTGAAGCTGGTGACAAATTGGAACAAGCT
GACAAGGACACCGTCACCAAGAAGGCTGAAGAGACTATTTCTTGGTTAGACAGCAACACC
ACTGCCAGCAAGGAAGAATTCGATGACAAGTTGAAGGAGTTGCAAGACATTGCCAACCCA
ATCATGTCTAAGTTGTACCAAGCTGGTGGTGCTCCAGGTGGCGCTGCAGGTGGTGCTCCA
GGCGGTTTCCCAGGTGGTGCTCCTCCAGCTCCAGAGGCTGAAGGTCCAACCGTTGAAGAA
GTTGATTAA
[0389] Further information on SSA1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000000004.
[0390] It will be appreciated that, by "SSA1", we include fragments
or variants thereof having equivalent SSA1-like activity.
[0391] SSA2 is another S. cerevisiae helper protein of interest for
the present invention. Ssa2p is an ATP binding protein that is
involved in protein folding and vacuolar import of proteins; member
of heat shock protein 70 (HSP70) family. It is associated with the
chaperonin-containing T-complex. It is present in the cytoplasm,
vacuolar membrane and cell wall. A published protein sequence for
the protein Ssa2p is as follows:
TABLE-US-00015 MSKAVGIDLGTTYSCVAHFSNDRVDIIANDQGNRTTPSFVGFTDTERLIG
DAAKNQAAMNPANTVFDAKRLIGRNFNDPEVQGDMKHFPFKLIDVDGKPQ
IQVEFKGETKNFTPEQISSMVLGKMKETAESYLGAKVNDAVVTVPAYFND
SQRQATKDAGTIAGLNVLRIINEPTAAAIAYGLDKKGKEEHVLIFDLGGG
TFDVSLLSIEDGIFEVKATAGDTHLGGEDFDNRLVNHFIQEFKRKNKKDL
STNQRALRRLRTACERAKRTLSSSAQTSVEIDSLFEGIDFYTSITRARFE
ELCADLFRSTLDPVEKVLRDAKLDKSQVDEIVLVGGSTRIPKVQKLVTDY
FNGKEPNRSINPDEAVAYGAAVQAAILTGDESSKTQDLLLLDVAPLSLGI
ETAGGVMTKLIPRNSTIPTKKSEVFSTYADNQPGVLIQVFEGERAKTKDN
NLLGKFELSGIPPAPRGVPQIEVTFDVDSNGILNVSAVEKGTGKSNKITI
TNDKGRLSKEDIEKMVAEAEKFKEEDEKESQRIASKNQLESIAYSLKNTI
SEAGDKLEQADKDAVTKKAEETIAWLDSNTTATKEEFDDQLKELQEVANP
IMSKLYQAGGAPEGAAPGGFPGGAPPAPEAEGPTVEEVD*
[0392] SSA2 is encoded by a non-essential gene comprising an ORF
that is 1.920 kbp in size and is located on chromosome XII. A
published nucleotide coding sequence of SSA2 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00016
ATGTCTAAAGCTGTCGGTATTGATTTAGGTACTACCTACTCCTGTGTTGCTCACTTCTCT
AATGATCGTGTTGACATTATTGCCAACGACCAAGGTAACAGAACCACTCCATCTTTCGTT
GGTTTCACTGATACTGAAAGATTGATTGGTGACGCTGCTAAGAACCAAGCTGCTATGAAC
CCAGCTAACACTGTTTTCGACGCTAAGCGTTTGATCGGTAGAAACTTCAATGACCCAGAA
GTCCAAGGTGATATGAAGCACTTCCCATTCAAGTTGATCGATGTTGACGGTAAGCCACAA
ATTCAAGTTGAATTTAAGGGTGAAACCAAGAACTTTACCCCAGAACAAATCTCCTCCATG
GTCTTGGGTAAGATGAAGGAAACTGCCGAATCTTACTTGGGTGCCAAGGTCAATGACGCT
GTCGTCACTGTCCCAGCTTACTTCAACGATTCTCAAAGACAAGCTACCAAGGATGCTGGT
ACCATTGCTGGTTTGAATGTCTTGCGTATTATTAACGAACCTACCGCCGCTGCCATTGCT
TACGGTTTGGACAAGAAGGGTAAGGAAGAACACGTCTTGATTTTCGACTTGGGTGGTGGT
ACTTTCGATGTCTCTTTGTTGTCCATTGAAGACGGTATCTTTGAAGTTAAGGCCACCGCT
GGTGACACCCATTTGGGTGGTGAAGATTTTGACAACAGATTGGTCAACCACTTCATCCAA
GAATTCAAGAGAAAGAACAAGAAGGACTTGTCTACCAACCAAAGAGCTTTGAGAAGATTA
AGAACTGCTTGTGAAAGAGCCAAGAGAACTTTGTCTTCCTCCGCTCAAACTTCCGTTGAA
ATTGACTCTTTGTTCGAAGGTATCGATTTCTACACTTCCATCACCAGAGCCAGATTCGAA
GAATTGTGTGCTGACTTGTTCAGATCTACTTTGGACCCAGTTGAAAAGGTCTTGAGAGAT
GCTAAATTGGATAAATCTCAAGTCGATGAAATTGTCTTGGTCGGTGGTTCTACCAGAATT
CCAAAGGTCCAAAAATTGGTCACTGACTACTTCAACGGTAAGGAACCAAACAGATCTATC
AACCCAGATGAAGCTGTTGCTTACGGTGCTGCTGTTCAAGCTGCTATTTTGACTGGTGAC
GAATCTTCCAAGACTCAAGATCTATTGTTGTTGGATGTCGCTCCATTATCCTTGGGTATT
GAAACTGCTGGTGGTGTCATGACCAAGTTGATTCCAAGAAACTCTACCATTCCAACTAAG
AAATCCGAAGTTTTCTCTACTTATGCTGACAACCAACCAGGTGTCTTGATTCAAGTCTTT
GAAGGTGAAAGAGCCAAGACTAAGGACAACAACTTGTTGGGTAAGTTCGAATTGAGTGGT
ATTCCACCAGCTCCAAGAGGTGTCCCACAAATTGAAGTCACTTTCGATGTCGACTCTAAC
GGTATTTTGAATGTTTCCGCCGTCGAAAAGGGTACTGGTAAGTCTAACAAGATCACTATT
ACCAACGACAAGGGTAGATTGTCCAAGGAAGATATCGAAAAGATGGTTGCTGAAGCCGAA
AAATTCAAGGAAGAAGATGAAAAGGAATCTCAAAGAATTGCTTCCAAGAACCAATTGGAA
TCCATTGCTTACTCTTTGAAGAACACCATTTCTGAAGCTGGTGACAAGCTAGAGCAAGCT
GACAAGGACGCTGTCACTAAGAAGGCTGAAGAAACTATTGCTTGGTTAGACAGCAACACC
ACTGCTACCAAGGAAGAATTCGATGACCAATTGAAGGAATTGCAAGAGGTTGCCAACCCA
ATCATGTCTAAATTGTACCAAGCTGGTGGTGCTCCAGAAGGCGCAGCTCCAGGTGGTTTC
CCAGGTGGTGCTCCTCCAGCTCCAGAAGCTGAAGGTCCAACTGTCGAAGAAGTTGATTAA
[0393] Further information on SSA2 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000003947.
[0394] It will be appreciated that, by "SSA2", we include fragments
or variants thereof having equivalent SSA2-like activity.
[0395] SSA3 is another S. cerevisiae helper protein of interest for
the present invention, which is also known as HSP70. Ssa3p is an
ATPase involved in protein folding and the response to stress. It
plays a role in SRP-dependent cotranslational protein-membrane
targeting and translocation and is a member of the heat shock
protein 70 (HSP70) family. SSA3 is localized to the cytoplasm. A
published protein sequence for the protein Ssa3p is as follows:
TABLE-US-00017 MSRAVGIDLGTTYSCVAHFSNDRVEIIANDQGNRTTPSYVAFTDTERLIG
DAAKNQAAINPHNTVFDAKRLIGRKFDDPEVTTDAKHFPFKVISRDGKPV
VQVEYKGETKTFTPEEISSMVLSKMKETAENYLGTTVNDAVVTVPAYFND
SQRQATKDAGTIAGMNVLRIINEPTAAAIAYGLDKKGRAEHNVLIFDLGG
GTFDVSLLSIDEGVFEVKATAGDTHLGGEDFDNRLVNHLATEFKRKTKKD
ISNNQRSLRRLRTAAERAKRALSSSSQTSIEIDSLFEGMDFYTSLTRARF
EELCADLFRSTLEPVEKVLKDSKLDKSQIDEIVLVGGSTRIPKIQKLVSD
FFNGKEPNRSINPDEAVAYGAAVQAAILTGDQSTKTQDLLLLDVAPLSLG
IETAGGIMTKLIPRNSTIPTKKSETFSTYADNQPGVLIQVFEGERTRTKD
NNLLGKFELSGIPPAPRGVPQIDVTFDIDANGILNVSALEKGTGKSNKIT
ITNDKGRLSKDDIDRMVSEAEKYRADDEREAERVQAKNQLESYAFTLKNT
INEASFKEKVGEDDAKRLETASQETIDWLDASQAASTDEYKDRQKELEGI
ANPIMTKFYGAGAGAGPGAGESGGFPGSMPNSGATGGGEDTGPTVEEVD*
[0396] SSA3 is encoded by a non-essential gene comprising an ORF
that is 1.950 kbp in size and is located on chromosome II. A
published nucleotide coding sequence of SSA3 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00018
ATGTCTAGAGCAGTTGGTATTGATTTGGGAACAACTTACTCGTGTGTTGCTCATTTTTCC
AATGATAGGGTAGAGATAATTGCAAATGATCAAGGTAATAGGACCACTCCATCGTATGTG
GCTTTCACAGACACCGAAAGATTAATTGGTGACGCCGCCAAAAATCAAGCTGCAATCAAT
CCTCATAATACAGTTTTTGATGCAAAGCGGTTAATTGGTCGTAAATTTGATGATCCTGAA
GTGACGACAGATGCCAAGCACTTCCCTTTCAAAGTTATATCCAGAGATGGTAAACCTGTA
GTGCAAGTAGAATATAAGGGTGAAACGAAAACATTTACGCCTGAGGAAATTTCTTCCATG
GTTTTAAGCAAAATGAAGGAAACTGCTGAGAACTATTTGGGAACTACGGTCAATGATGCT
GTTGTAACTGTTCCTGCATATTTCAATGATTCTCAAAGACAAGCCACTAAGGATGCAGGA
ACTATTGCAGGGATGAACGTTTTACGTATTATCAATGAACCCACTGCAGCAGCAATTGCT
TATGGCTTGGATAAGAAAGGCAGGGCTGAGCACAATGTCCTGATTTTTGATTTGGGTGGT
GGTACTTTTGACGTCTCTTTACTTTCAATTGATGAGGGTGTTTTTGAGGTTAAGGCTACC
GCAGGAGACACTCATTTAGGTGGTGAAGATTTTGATAATAGGTTGGTGAACCATTTAGCC
ACTGAATTCAAAAGGAAAACGAAAAAGGACATCTCTAATAATCAAAGATCGTTAAGAAGA
TTGAGAACTGCGGCAGAAAGAGCTAAGAGAGCGCTTTCTTCCTCATCTCAAACCTCGATC
GAGATCGATTCTTTATTTGAAGGTATGGATTTCTACACTTCGTTAACAAGGGCAAGGTTT
GAAGAGCTATGTGCTGATTTATTCAGATCCACATTGGAACCAGTAGAAAAGGTTCTTAAA
GATTCGAAGCTGGACAAGTCCCAAATTGATGAGATTGTGTTAGTCGGTGGATCTACCAGA
ATCCCAAAGATTCAGAAATTAGTTTCTGACTTCTTCAATGGCAAAGAGCCTAATCGTTCT
ATCAACCCGGATGAGGCTGTTGCTTATGGTGCAGCCGTTCAAGCTGCCATTTTAACCGGC
GATCAATCAACAAAGACACAAGATTTACTATTATTGGATGTTGCGCCATTGTCCCTAGGA
ATTGAAACTGCAGGCGGCATAATGACTAAGCTAATTCCTAGAAACTCAACGATTCCAACA
AAGAAATCGGAAACCTTCTCTACCTATGCAGATAATCAACCTGGTGTTTTAATTCAAGTC
TTTGAAGGTGAAAGAACAAGAACAAAGGATAATAACTTACTTGGTAAATTCGAATTAAGT
GGCATTCCGCCTGCTCCCAGAGGTGTGCCTCAAATTGATGTTACCTTTGATATCGACGCT
AATGGTATTCTTAATGTGTCTGCTTTGGAAAAGGGTACTGGTAAGAGTAACAAAATCACG
ATCACTAACGATAAAGGTAGGCTCTCGAAGGATGATATTGATAGGATGGTTTCTGAAGCT
GAAAAATATAGGGCTGACGATGAAAGGGAGGCAGAACGAGTTCAGGCTAAGAATCAGCTT
GAATCGTATGCATTTACTTTGAAGAATACCATAAACGAAGCAAGTTTCAAAGAGAAAGTA
GGTGAAGATGATGCAAAGAGATTAGAAACAGCGTCTCAGGAAACCATTGACTGGTTAGAT
GCATCGCAGGCAGCCTCTACGGACGAATATAAGGATAGACAAAAGGAGTTGGAAGGCATT
GCCAATCCAATAATGACGAAATTTTACGGTGCTGGTGCCGGCGCAGGTCCTGGAGCGGGG
GAATCCGGTGGATTCCCCGGATCCATGCCCAACTCGGGTGCTACGGGAGGTGGAGAAGAT
ACAGGTCCAACAGTGGAAGAGGTTGATTGA
[0397] Further information on SSA3 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000000171.
[0398] It will be appreciated that, by "SSA3", we include fragments
or variants thereof having equivalent SSA3-like activity.
[0399] SSA4 is another S. cerevisiae helper protein of interest for
the present invention. Ssa4p is a heat shock protein that is highly
induced upon stress. It plays a role in SRP-dependent
cotranslational protein-membrane targeting and translocation;
member of the HSP70 family. It is a cytoplasmic protein that
concentrates in nuclei upon starvation. A published protein
sequence for the protein Ssa4p is as follows:
TABLE-US-00019 MSKAVGIDLGTTYSCVAHFANDRVEIIANDQGNRTTPSYVAFTDTERLIG
DAAKNQAAMNPHNTVFDAKRLIGRKFDDPEVTNDAKHYPFKVIDKGGKPV
VQVEYKGETKTFTPEEISSMILTKMKETAENFLGTEVKDAVVTVPAYFND
SQRQATKDAGTIAGLNVLRIINEPTAAAIAYGLDKKSQKEHNVLIFDLGG
GTFDVSLLSIDEGVFEVKATAGDTHLGGEDFDSRLVNFLAEEFKRKNKKD
LTTNQRSLRRLRTAAERAKRTLSSSAQTSIEIDSLFEGIDFYTSITRARF
EELCADLFRSTLEPVEKVLADSKLDKSQIDEIVLVGGSTRIPKVQKLVSD
FFNGKEPNRSINPDEAVAYGAAVQAAILTGDQSSTTQDLLLLDVAPLSLG
IETAGGIMTKLIPRNSTIPTKKSEVFSTYADNQPGVLIQVFEGERTRTKD
NNLLGKFELSGIPPAPRGVPQIEVTFDIDANGILNVSAVEKGTGKSNKIT
ITNDKGRLSKEDIDKMVAEAEKFKAEDEQEAQRVQAKNQLESYAFTLKNS
VSENNFKEKVGEEDARKLEAAAQDAINWLDASQAASTEEYKERQKELEGV
ANPIMSKFYGAAGGAPGAGPVPGAGAGPTGAPDNGPTVEEVD*
[0400] SSA4 is encoded by a non-essential gene comprising an ORF
that is 1.929 kbp in size and is located on chromosome V. A
published nucleotide coding sequence of SSA4 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00020
ATGTCAAAAGCTGTTGGTATTGATTTAGGTACAACCTATTCATGTGTTGCTCATTTTGCA
AACGATAGGGTTGAAATTATCGCTAACGATCAAGGTAATAGAACGACGCCTTCTTATGTG
GCTTTTACTGACACAGAAAGGCTAATTGGTGACGCTGCGAAGAATCAAGCTGCGATGAAC
CCACATAATACAGTATTCGATGCTAAGCGTCTGATCGGACGTAAATTCGATGATCCAGAA
GTGACGAACGATGCTAAGCATTACCCATTCAAAGTGATTGACAAGGGAGGTAAACCGGTA
GTGCAAGTGGAATATAAAGGCGAGACAAAGACATTTACTCCAGAAGAAATTTCCTCAATG
ATCTTGACAAAGATGAAGGAGACTGCTGAGAACTTTTTAGGAACAGAAGTGAAAGATGCT
GTAGTAACGGTTCCAGCCTATTTCAACGATTCACAAAGGCAAGCAACAAAAGATGCCGGT
ACAATCGCGGGCTTGAACGTTCTTCGTATCATTAATGAACCTACAGCTGCCGCTATTGCG
TATGGGCTGGACAAGAAATCGCAGAAGGAGCACAACGTCTTGATCTTTGATTTAGGTGGT
GGTACTTTTGATGTCTCTCTGCTATCCATAGATGAAGGTGTCTTTGAGGTTAAGGCTACT
GCTGGTGACACTCACTTGGGTGGTGAAGATTTCGATAGTAGGCTGGTTAACTTTCTAGCC
GAGGAGTTCAAAAGAAAAAATAAAAAGGATCTAACAACTAACCAAAGGTCCCTAAGGAGG
TTAAGGACCGCCGCTGAAAGGGCCAAGAGAACTCTGTCTTCGTCTGCTCAGACATCTATA
GAAATAGATTCATTATTTGAGGGTATCGATTTCTATACTTCCATTACAAGGGCAAGATTT
GAAGAATTATGTGCTGATTTGTTTAGATCTACATTGGAGCCAGTGGAAAAAGTTTTGGCT
GATTCAAAATTAGATAAGTCACAAATTGATGAAATTGTACTTGTTGGTGGTTCAACAAGA
ATTCCAAAAGTACAAAAACTGGTTTCTGATTTTTTCAATGGTAAAGAACCAAACCGTTCG
ATTAACCCTGATGAGGCCGTCGCTTATGGTGCTGCCGTACAGGCTGCCATCTTAACGGGT
GACCAGTCGTCGACGACCCAAGATTTACTGTTGCTGGATGTTGCACCATTATCTCTAGGT
ATTGAAACTGCAGGTGGTATTATGACAAAGTTGATCCCAAGAAATTCGACTATCCCAACA
AAAAAATCGGAAGTGTTTTCCACCTACGCTGACAACCAACCTGGTGTGTTGATACAAGTT
TTTGAGGGTGAAAGGACAAGGACAAAAGACAACAATCTACTGGGTAAATTTGAGTTGAGC
GGTATTCCACCCGCTCCAAGAGGCGTACCACAAATTGAAGTTACATTTGATATCGATGCA
AATGGTATTCTGAACGTATCTGCCGTTGAAAAAGGTACTGGTAAATCTAACAAGATTACA
ATTACTAACGATAAGGGAAGATTATCGAAGGAAGATATCGATAAAATGGTTGCTGAGGCA
GAAAAGTTCAAGGCCGAAGATGAACAAGAAGCTCAACGTGTTCAAGCTAAGAATCAGCTA
GAATCGTACGCGTTTACTTTGAAAAATTCTGTGAGCGAAAATAACTTCAAGGAGAAGGTG
GGTGAAGAGGATGCCAGGAAATTGGAAGCCGCCGCCCAAGATGCTATAAATTGGTTAGAT
GCTTCGCAAGCGGCCTCCACCGAGGAATACAAGGAAAGGCAAAAGGAACTAGAAGGTGTT
GCAAACCCCATTATGAGTAAATTTTACGGAGCTGCAGGTGGTGCCCCAGGAGCAGGCCCA
GTTCCGGGTGCTGGAGCAGGCCCCACTGGAGCACCAGACAACGGCCCAACGGTTGAAGAG
GTTGATTAG
[0401] Further information on SSA4 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000000905.
[0402] It will be appreciated that, by "SSA4", we include fragments
or variants thereof having equivalent SSA4-like activity.
[0403] SSE1 is another S. cerevisiae helper protein of interest for
the present invention and is also known as LPG3 and MSI3. Sse1p is
an ATPase that is a component of the heat shock protein Hsp90
chaperone complex. It binds unfolded proteins and is a member of
the heat shock protein 70 (HSP70) family. It is localized to the
cytoplasm. A published protein sequence for the protein Sse1p is as
follows:
TABLE-US-00021
MSTPFGLDLGNNNSVLAVARNRGIDIVVNEVSNRSTPSVVGFGPKNRYLGETGKNKQTSN
IKNTVANLKRIIGLDYHHPDFEQESKHFTSKLVELDDKKTGAEVRFAGEKHVFSATQLAA
MFIDKVKDTVKQDTKANITDVCIAVPPWYTEEQRYNIADAARIAGLNPVRIVNDVTAAGV
SYGIFKTDLPEGEEKPRIVAFVDIGHSSYTCSIMAFKKGQLKVLGTACDKHFGGRDFDLA
ITEHFADEFKTKYKIDIRENPKAYNRILTAAEKLKKVLSANTNAPFSVESVMNDVDVSSQ
LSREELEELVKPLLERVTEPVTKALAQAKLSAEEVDFVEIIGGTTRIPTLKQSISEAFGK
PLSTTLNQDEAIAKGAAFICAIHSPTLRVRPFKFEDIHPYSVSYSWDKQVEDEDHMEVFP
AGSSFPSTKLITLNRTGDFSMAASYTDITQLPPNTPEQIANWEITGVQLPEGQDSVPVKL
KLRCDPSGLHTIEEAYTIEDIEVEEPIPLPEDAPEDAEQEFKKVTKTVKKDDLTIVAHTF
GLDAKKLNELIEKENEMLAQDKLVAETEDRKNTLEEYIYTLRGKLEEEYAPFASDAEKTK
LQGMLNKAEEWLYDEGFDSIKAKYIAKYEELASLGNIIRGRYLAKEEEKKQAIRSKQEAS
QMAAMAEKLAAQRKAEAEKKEEKKDTEGDVDMD*
[0404] SSE1 is encoded by a non-essential gene comprising an ORF
that is 2.082 kbp in size and is located on chromosome XVI. A
published nucleotide coding sequence of SSE1 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00022
ATGAGTACTCCATTTGGTTTAGATTTAGGTAACAATAACTCTGTCCTTGCCGTTGCTAGA
AACAGAGGTATCGACATTGTCGTTAATGAAGTCTCTAACCGTTCCACCCCATCTGTTGTT
GGTTTTGGTCCAAAGAACAGATACTTGGGTGAAACTGGTAAGAACAAGCAGACTTCCAAC
ATCAAGAACACTGTCGCCAACTTGAAAAGAATTATTGGTTTGGATTACCACCATCCAGAT
TTCGAGCAAGAATCTAAGCACTTCACCTCTAAGTTGGTTGAATTGGATGACAAGAAGACT
GGTGCCGAAGTTAGATTCGCTGGTGAGAAACATGTTTTTTCAGCTACTCAACTAGCTGCC
ATGTTCATCGACAAAGTCAAGGACACCGTCAAGCAGGACACAAAGGCAAATATTACCGAT
GTTTGTATTGCTGTCCCACCTTGGTACACCGAAGAACAACGTTACAACATTGCTGATGCT
GCTAGAATTGCTGGTTTGAACCCTGTTAGAATTGTCAACGACGTTACTGCTGCCGGTGTT
TCTTACGGTATCTTCAAGACTGATTTGCCTGAAGGCGAAGAAAAGCCAAGAATTGTTGCC
TTTGTTGATATTGGTCACTCTTCCTACACCTGTTCTATCATGGCCTTCAAGAAGGGTCAA
TTGAAAGTCTTAGGAACTGCCTGCGACAAGCATTTTGGTGGTAGGGACTTCGATTTGGCT
ATAACAGAACATTTCGCCGATGAGTTCAAAACTAAATACAAGATTGACATCAGAGAAAAT
CCAAAGGCTTACAACAGAATTCTAACTGCTGCTGAAAAGTTGAAGAAAGTTTTGTCTGCT
AATACTAATGCCCCATTCTCTGTTGAATCCGTCATGAACGACGTTGATGTTTCCTCTCAA
TTATCTCGTGAAGAATTAGAAGAATTGGTCAAGCCATTGTTGGAACGTGTTACTGAACCA
GTTACCAAAGCTTTAGCTCAAGCCAAATTATCTGCTGAAGAAGTTGATTTTGTTGAAATT
ATTGGTGGTACTACTCGTATCCCAACATTGAAACAATCCATTTCTGAAGCCTTCGGCAAG
CCATTGTCCACCACTTTGAACCAAGATGAAGCCATCGCCAAGGGTGCCGCCTTTATTTGC
GCCATTCACTCTCCAACTCTAAGAGTTAGACCATTCAAGTTTGAGGATATCCATCCTTAC
TCTGTCTCTTACTCTTGGGACAAGCAAGTTGAGGACGAAGACCACATGGAAGTTTTCCCA
GCTGGTTCATCCTTCCCATCTACTAAATTGATCACTTTGAACCGTACGGGTGACTTTTCA
ATGGCTGCTAGCTACACTGACATCACACAGTTACCACCAAACACTCCAGAACAAATCGCT
AACTGGGAGATCACTGGTGTTCAATTACCAGAAGGTCAAGACTCTGTTCCTGTTAAGTTA
AAGTTGAGATGCGACCCCTCTGGTTTACACACAATTGAAGAGGCTTACACTATTGAAGAT
ATTGAAGTTGAAGAACCTATTCCATTACCAGAAGATGCTCCAGAAGATGCTGAGCAAGAA
TTTAAGAAGGTTACTAAAACTGTAAAGAAGGATGACTTAACCATCGTTGCACACACCTTT
GGCCTAGACGCTAAAAAGTTGAATGAATTAATTGAAAAAGAAAATGAAATGCTTGCTCAA
GATAAGCTAGTTGCTGAGACAGAAGACCGTAAGAACACTCTTGAAGAGTACATCTACACA
TTGCGTGGTAAGTTGGAAGAAGAGTATGCTCCATTTGCTTCCGATGCTGAAAAGACGAAG
TTACAAGGTATGTTAAACAAGGCCGAAGAGTGGTTATACGATGAAGGTTTCGATTCCATC
AAAGCTAAGTACATTGCCAAATACGAAGAATTGGCTTCTCTAGGTAACATTATTAGAGGT
AGATACTTGGCTAAAGAAGAAGAAAAGAAGCAAGCTATAAGATCTAAGCAAGAAGCATCC
CAAATGGCTGCTATGGCTGAAAAGTTGGCTGCTCAAAGAAAGGCAGAAGCTGAAAAGAAG
GAAGAAAAGAAGGACACTGAAGGTGATGTTGACATGGACTAA
[0405] Further information on SSE1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000006027.
[0406] It will be appreciated that, by "SSE1", we include fragments
or variants thereof having equivalent SSE1-like activity.
[0407] SSE2 is another S. cerevisiae helper protein of interest for
the present invention. Sse2p is a member of the heat shock protein
70 (HSP70) family. It may be involved in protein folding and is
localised to the cytoplasm. It is highly homologous to the heat
shock protein Sse1p. A published protein sequence for the protein
Sse2p is as follows:
TABLE-US-00023
MSTPFGLDLGNNNSVLAVARNRGIDVVVNEVSNRSTPSLVGFGPRNRYLGESGKTKQTSN
VKNTVENLKRIIGLKFKDPEFDIENKFFTSKLVQLKNGKVGVEVEFGGKTHVFSATQLTA
MFIDKVKHTVQEETKSSITDVCLAVPVWYSEEQRYNIADAARIAGLNPVRIVNDVTAAAV
SYGVFKNDLPGPEEKPRIIGLVDIGHSTYTCSIMAFRKGEMKVLGTAYDKHFGGRDFDRA
ITEHFADQFKDKYKIDIRKNPKAYNRILIAAEKLKKVLSANTTAPFSVESVMDDIDVSSQ
LSREELEELVEPLLKRVTYPITNALAQAKLTVNDIDFVEIIGGTTRIPVLKKSISDVFGK
PLSSTLNQDEAVAKGAAFICAIHSPTLRVRPFKFEDIDPYSVSYTWDKQVDDEDRLEVFP
ANSSYPSTKLITLHRTGDFSMKAVYTHPSKLPKGTSTTIAKWSFTGVKVPKDQDFIPVKV
KLRCDPSGLHIIENAYTTEDITVQEPVPLPEDAPEDAEPQFKEVTKTIKKDVLGMTAKTF
ALNPVELNDLIEKENELRNQDKLVAETEDRKNALEEYIYTLRAKLDDEYSDFASDAEKEK
LKNMLATTENWLYGDGDDSTKAKYIAKYEELASLGNIIRGRYLAKEEEKRQALRANQETS
KMNDIAEKLAEQRRARAASDDSDDNNDENMDLD*
[0408] SSE2 is encoded by a non-essential gene comprising an ORF
that is 2.082 kbp in size and is located on chromosome II. A
published nucleotide coding sequence of SSE2 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00024
ATGAGCACTCCATTTGGCTTAGATTTAGGTAACAATAACTCAGTACTAGCAGTTGCCAGA
AATAGGGGTATTGATGTCGTTGTCAATGAAGTTTCTAATAGGTCTACACCATCCTTGGTC
GGCTTTGGCCCCAGAAATAGGTACTTAGGTGAATCTGGTAAAACTAAGCAAACATCGAAT
GTTAAAAACACTGTGGAAAACTTGAAAAGAATCATTGGACTAAAGTTCAAAGACCCTGAA
TTTGATATCGAGAATAAGTTCTTCACTTCGAAATTGGTACAGCTAAAAAATGGTAAAGTT
GGTGTGGAAGTGGAGTTCGGCGGTAAAACACACGTATTTTCAGCTACTCAACTGACTGCT
ATGTTCATTGATAAGGTGAAGCACACCGTTCAAGAGGAAACGAAGTCATCAATTACCGAT
GTCTGCCTCGCAGTTCCTGTATGGTATTCGGAAGAACAACGTTATAACATAGCCGATGCT
GCCAGAATTGCAGGATTAAATCCTGTAAGGATTGTCAACGATGTGACTGCAGCCGCCGTT
TCGTACGGCGTCTTCAAGAATGATCTGCCAGGTCCTGAAGAAAAGCCAAGAATCATTGGC
TTAGTGGACATTGGGCATTCTACCTACACCTGTTCTATTATGGCTTTCCGCAAAGGCGAA
ATGAAAGTATTAGGTACTGCTTATGACAAGCACTTTGGTGGTAGAGATTTCGATCGCGCA
ATCACAGAACATTTTGCTGATCAGTTTAAGGACAAGTACAAGATTGACATTAGGAAAAAT
CCGAAAGCTTATAACAGAATTTTAATCGCTGCTGAAAAATTAAATAAAGTGCTTTCTGCG
AACACTACTGCCCCCTTCTCCGTTGAATCTGTTATGGATGATATCGACGTTTCCTCTCAA
TTGAGCCGTGAAGAGCTGGAAGAATTAGTAGAGCCCTTGTTGAAGCGTGTGACGTATCCA
ATCACCAATGCATTGGCTCAAGCTAAATTAACTGTCAATGATATTGACTTCGTAGAAATA
ATTGGTGGTACAACCCGTATCCCAGTTTTAAAGAAGTCAATTTCTGATGTTTTTGGAAAA
CCTTTGTCATCTACTTTAAATCAAGACGAAGCTGTGGCCAAGGGGGCCGCTTTCATATGT
GCCATTCACTCTCCAACTTTAAGGGTCAGGCCGTTTAAATTTGAAGATATTGATCCGTAT
TCAGTGTCATACACTTGGGATAAGCAGGTCGATGACGAAGACCGTTTGGAAGTATTCCCT
GCTAATTCATCATATCCATCAACTAAACTAATTACTTTACATCGTACTGGAGATTTCAGC
ATGAAAGCGGTGTACACTCATCCTTCGAAACTGCCAAAAGGTACTTCCACCACTATTGCA
AAATGGAGCTTCACTGGGGTCAAGGTTCCTAAAGATCAAGATTTTATTCCTGTAAAGGTC
AAGTTAAGATGCGATCCTTCCGGCTTGCATATTATCGAGAACGCTTACACAACGGAAGAT
ATTACGGTTCAAGAGCCAGTGCCTTTACCGGAAGACGCACCAGAAGATGCCGAGCCCCAG
TTTAAAGAAGTTACTAAAACAATTAAGAAAGATGTGCTAGGTATGACTGCAAAAACATTC
GCGCTAAACCCGGTTGAGTTGAACGATCTAATTGAAAAAGAGAATGAATTAAGAAACCAG
GATAAGTTAGTTGCCGAAACCGAGGATCGCAAAAATGCCCTTGAAGAGTATATTTATACC
CTTCGTGCCAAACTCGATGATGAATACTCCGATTTTGCGTCTGACGCAGAAAAAGAAAAG
CTAAAAAACATGTTAGCCACTACTGAAAATTGGTTATATGGTGATGGTGACGATTCTACC
AAGGCAAAATACATTGCTAAATATGAGGAGCTGGCATCGTTGGGGAATATTATTAGAGGT
AGATATTTAGCAAAGGAGGAAGAAAAAAGACAAGCACTCAGAGCGAATCAAGAAACTTCT
AAAATGAATGATATTGCTGAAAAATTGGCTGAGCAAAGAAGGGCACGCGCTGCAAGTGAT
GATAGCGATGACAACAATGATGAAAACATGGACCTTGATTAA
[0409] Further information on SSE2 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000000373.
[0410] It will be appreciated that, by "SSE2", we include fragments
or variants thereof having equivalent SSE2-like activity.
[0411] SSB1 is another S. cerevisiae helper protein of interest for
the present invention and is also known as YG101. Ssb1p is a
cytoplasmic ATPase that is a ribosome-associated molecular
chaperone. It may be involved in the folding of newly-synthesized
polypeptide chains and is a member of the heat shock protein 70
(HSP70) family. It interacts with the phosphatase subunit Reg1p. A
published protein sequence for the protein Ssb1p is as follows:
TABLE-US-00025
MAEGVFQGAIGIDLGTTYSCVATYESSVEIIANEQGNRVTPSFVAFTPEERLIGDAAKNQ
AALNPRNTVFDAKRLIGRRFDDESVQKDMKTWPFKVIDVDGNPVIEVQYLEETKTFSPQE
ISAMVLTKMKEIAEAKIGKKVEKAVITVPAYFNDAQRQATKDAGAISGLNVLRIINEPTA
AAIAYGLGAGKSEKERHVLIFDLGGGTFDVSLLHIAGGVYTVKSTSGNTHLGGQDFDTNL
LEHFKAEFKKKTGLDISDDARALRRLRTAAERAKRTLSSVTQTTVEVDSLFDGEDFESSL
TRARFEDLNAALFKSTLEPVEQVLKDAKISKSQIDEVVLVGGSTRIPKVQKLLSDFFDGK
QLEKSINPDEAVAYGAAVQGAILTGQSTSDETKDLLLLDVAPLSLGVGMQGDMFGIVVPR
NTTVPTIKRRTFTTCADNQTTVQFPVYQGERVNCKENTLLGEFDLKNIPMMPAGEPVLEA
IFEVDANGILKVTAVEKSTGKSSNITISNAVGRLSSEEIEKMVNQAEEFKAADEAFAKKH
EARQRLESYVASIEQTVTDPVLSSKLKRGSKSKIEAALSDALAALQIEDPSADELRKAEV
GLKRVVTKAMSSR*
[0412] SSB1 is encoded by a non-essential gene comprising an ORF
that is 1.842 kbp in size and is located on chromosome IV. A
published nucleotide coding sequence of SSB1 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00026
ATGGCTGAAGGTGTTTTCCAAGGTGCTATCGGTATCGATTTAGGTACAACCTACTCTTGT
GTTGCTACTTACGAATCCTCCGTTGAAATTATTGCCAACGAACAAGGTAACAGAGTCACC
CCATCTTTCGTTGCTTTCACTCCAGAAGAAAGATTGATTGGTGATGCTGCCAAGAACCAA
GCTGCTTTGAACCCAAGAAACACTGTCTTCGATGCTAAGCGTTTGATTGGTAGAAGATTC
GACGACGAATCTGTTCAAAAGGACATGAAGACCTGGCCTTTCAAGGTTATCGACGTCGAT
GGTAACCCAGTCATCGAAGTCCAATACTTGGAAGAAACCAAGACTTTCTCCCCACAAGAA
ATTTCCGCTATGGTTTTGACCAAGATGAAGGAAATTGCTGAAGCTAAGATTGGTAAGAAG
GTTGAAAAGGCCGTCATTACTGTCCCAGCTTACTTTAACGACGCTCAAAGACAAGCTACC
AAGGATGCCGGTGCCATTTCTGGTTTGAACGTTTTGCGTATCATCAACGAACCTACTGCC
GCTGCTATTGCTTACGGTCTAGGTGCTGGTAAGTCCGAAAAGGAAAGACATGTTTTGATT
TTCGATTTGGGTGGTGGTACTTTCGATGTTTCCTTGTTGCACATTGCTGGTGGTGTTTAC
ACTGTTAAATCTACTTCCGGTAACACTCACTTGGGTGGTCAAGATTTCGACACCAACTTG
TTGGAACACTTCAAGGCTGAATTCAAGAAGAAGACTGGTTTGGACATCTCCGACGATGCC
AGAGCTTTGAGAAGATTGAGAACTGCTGCTGAAAGAGCTAAGAGAACCTTATCTTCTGTC
ACTCAAACTACCGTTGAAGTTGACTCTTTGTTTGACGGTGAAGATTTCGAATCCTCTTTG
ACTAGAGCTAGATTTGAAGACTTGAACGCCGCATTGTTCAAGTCTACTTTGGAACCTGTT
GAACAAGTTTTGAAGGATGCTAAGATCTCTAAGTCTCAAATCGACGAAGTTGTCTTGGTT
GGTGGTTCCACCAGAATTCCAAAGGTCCAAAAGTTGTTGTCTGACTTCTTTGACGGTAAG
CAATTGGAAAAATCTATTAACCCAGATGAAGCTGTTGCTTACGGTGCTGCTGTTCAAGGT
GCTATCTTGACCGGCCAATCCACATCTGACGAAACCAAGGACTTGTTGTTGTTAGATGTT
GCTCCATTATCTCTAGGTGTTGGTATGCAAGGTGACATGTTCGGTATCGTTGTTCCAAGA
AACACTACTGTTCCAACCATCAAGAGAAGAACCTTTACTACATGTGCTGACAACCAAACC
ACCGTTCAATTCCCAGTCTACCAAGGTGAACGTGTTAACTGTAAAGAAAACACTTTGTTG
GGTGAATTCGACTTGAAGAACATCCCAATGATGCCAGCTGGTGAACCAGTCTTGGAAGCT
ATCTTCGAAGTTGATGCTAACGGTATCTTGAAGGTTACTGCCGTCGAAAAGTCTACCGGT
AAGTCTTCTAACATCACTATCTCTAACGCTGTTGGTAGATTGTCTTCTGAAGAAATTGAA
AAGATGGTTAACCAAGCTGAAGAGTTCAAGGCTGCCGATGAAGCTTTTGCCAAGAAGCAC
GAAGCTAGACAAAGATTGGAATCCTACGTTGCCTCCATCGAACAAACTGTCACTGACCCA
GTCTTGTCTTCTAAATTGAAGAGAGGTTCCAAGTCCAAGATTGAAGCTGCTTTGTCCGAT
GCTTTGGCTGCTTTGCAAATCGAAGACCCATCTGCTGATGAATTGAGAAAGGCTGAAGTT
GGTTTGAAGAGAGTTGTCACCAAGGCCATGTCTTCTCGTTAA
[0413] Further information on SSB1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000002388.
[0414] It will be appreciated that, by "SSB1", we include fragments
or variants thereof having equivalent SSB1-like activity.
[0415] SSB2 is another S. cerevisiae helper protein of interest for
the present invention. Ssb2p is a cytoplasmic ATPase that is a
ribosome-associated molecular chaperone. It may be involved in the
folding of newly-synthesized polypeptide chains. It is a member of
the heat shock protein 70 (HSP70) family and is a homolog of SSB1.
A published protein sequence for the protein Ssb2p is as
follows:
TABLE-US-00027
MAEGVFQGAIGIDLGTTYSCVATYESSVEIIANEQGNRVTPSFVAFTPQERLIGDAAKNQ
AALNPRNTVFDAKRLIGRRFDDESVQKDMKTWPFKVIDVDGNPVIEVQYLEETKTFSPQE
ISAMVLTKMKEIAEAKIGKKVEKAVITVPAYFNDAQRQATKDAGAISGLNVLRIINEPTA
AAIAYGLGAGKSEKERHVLIFDLGGGTFDVSLLHIAGGVYTVKSTSGNTHLGGQDFDTNL
LEHFKAEFKKKTGLDISDDARALRRLRTAAERAKRTLSSVTQTTVEVDSLFDGEDFESSL
TRARFEDLNAALFKSTLEPVEQVLKDAKISKSQIDEVVLVGGSTRIPKVQKLLSDFFDGK
QLEKSINPDEAVAYGAAVQGAILTGQSTSDETKDLLLLDVAPLSLGVGMQGDIFGIVVPR
NTTVPTIKRRTFTTVSDNQTTVQFPVYQGERVNCKENTLLGEFDLKNIPMMPAGEPVLEA
IFEVDANGILKVTAVEKSTGKSSNITISNAVGRLSSEEIEKMVNQAEEFKAADEAFAKKH
EARQRLESYVASIEQTVTDPVLSSKLKRGSKSKIEAALSDALAALQIEDPSADELRKAEV
GLKRVVTKAMSSR*
[0416] SSB2 is encoded by a non-essential gene comprising an ORF
that is 1.842 kbp in size and is located on chromosome XIV. A
published nucleotide coding sequence of SSB2 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00028
ATGGCTGAAGGTGTTTTCCAAGGTGCTATCGGTATCGATTTAGGTACAACATACTCTTGT
GTTGCTACTTATGAATCTTCCGTTGAAATTATTGCCAACGAACAAGGTAACAGAGTTACT
CCATCTTTCGTTGCCTTCACCCCACAGGAAAGATTGATCGGTGATGCTGCCAAGAACCAA
GCTGCTTTGAACCCAAGAAACACTGTTTTTGATGCTAAGCGTTTGATTGGTAGAAGATTC
GACGACGAGTCTGTCCAAAAGGACATGAAGACCTGGCCTTTCAAGGTTATCGACGTCGAT
GGTAACCCAGTCATTGAAGTCCAATACTTGGAAGAAACCAAGACTTTCTCCCCACAAGAA
ATTTCCGCTATGGTCTTGACCAAGATGAAGGAAATTGCTGAAGCTAAGATTGGTAAGAAG
GTTGAAAAGGCTGTCATTACTGTCCCAGCTTACTTTAACGATGCCCAAAGACAAGCTACC
AAGGATGCCGGTGCCATTTCTGGTTTGAACGTTTTGCGTATCATCAACGAACCTACTGCC
GCTGCTATTGCTTACGGTCTAGGTGCTGGTAAGTCCGAAAAGGAAAGACATGTTTTGATT
TTCGATTTGGGTGGTGGTACTTTCGATGTTTCCTTGTTGCACATTGCTGGTGGTGTTTAC
ACTGTTAAATCTACTTCCGGTAACACTCACTTGGGTGGTCAAGATTTCGACACCAACTTG
TTGGAACACTTCAAGGCTGAATTCAAGAAGAAGACTGGTTTGGACATCTCCGACGATGCC
AGAGCTTTGAGAAGATTGAGAACTGCTGCTGAAAGAGCTAAGAGAACCTTATCTTCTGTC
ACTCAAACTACCGTTGAAGTTGACTCTTTGTTTGACGGTGAAGATTTCGAATCCTCTTTG
ACTAGAGCTAGATTTGAAGACTTGAACGCCGCATTGTTCAAGTCTACTTTGGAACCTGTT
GAACAAGTTTTGAAGGATGCTAAGATCTCTAAGTCTCAAATCGACGAAGTTGTCTTGGTT
GGTGGTTCTACCAGAATTCCAAAGGTCCAAAAGTTGTTGTCTGACTTCTTTGACGGTAAG
CAATTGGAAAAATCTATTAACCCAGATGAAGCTGTTGCTTACGGTGCTGCTGTTCAAGGT
GCTATCTTGACTGGCCAATCCACATCTGACGAAACCAAGGACTTGTTGTTGTTAGATGTT
GCTCCATTATCTCTAGGTGTTGGTATGCAAGGTGACATTTTCGGTATTGTTGTCCCAAGA
AACACAACTGTTCCAACCATCAAGAGAAGAACCTTCACAACTGTCAGTGACAACCAAACC
ACCGTTCAATTCCCAGTCTACCAAGGTGAACGTGTCAACTGTAAAGAAAACACTTTGTTG
GGTGAATTCGACTTGAAGAACATCCCAATGATGCCAGCTGGTGAACCAGTCTTGGAAGCT
ATCTTCGAAGTTGATGCTAACGGTATCTTGAAGGTTACTGCCGTCGAAAAGTCTACCGGT
AAGTCTTCTAACATCACTATCTCCAACGCTGTCGGTAGATTGTCTTCTGAAGAAATTGAA
AAGATGGTTAACCAAGCCGAAGAGTTCAAGGCTGCTGATGAAGCTTTTGCTAAGAAGCAC
GAAGCTAGACAAAGACTAGAATCCTACGTCGCTTCCATCGAACAAACCGTCACTGACCCA
GTCTTGTCTTCTAAATTGAAGAGAGGTTCCAAGTCCAAGATCGAAGCTGCTTTGTCCGAT
GCTTTGGCTGCTTTGCAAATCGAAGACCCATCCGCTGATGAGTTGAGAAAGGCAGAAGTT
GGTTTGAAGAGAGTTGTCACCAAGGCCATGTCTTCTCGTTAA
[0417] Further information on SSB2 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000005153.
[0418] It will be appreciated that, by "SSB2", we include fragments
or variants thereof having equivalent SSB2-like activity.
[0419] ECM10 is another S. cerevisiae helper protein of interest
for the present invention and is also known as SSC3. Ecm10p is a
heat shock protein of the Hsp70 family, which is localised in
mitochondrial nucleoids. It is thought to play a role in protein
translocation. It interacts with Mge1p in an ATP-dependent manner.
Over-expression has been shown to induce extensive mitochondrial
DNA aggregations. A published protein sequence for the protein
Ecm10p is as follows:
TABLE-US-00029
MLPSWKAFKAHNILRILTRFQSTKIPDAVIGIDLGTTNSAVAIMEGKVPRIIENAEGSRT
TPSVVAFTKDGERLVGEPAKRQSVINSENTLFATKRLIGRRFEDAEVQRDINQVPFKIVK
HSNGDAWVEARNRTYSPAQIGGFILNKMKETAEAYLAKSVKNAVVTVPAYFNDAQRQATK
DAGQIIGLNVLRVVNEPTAAALAYGLDKSEPKVIAVFDLGGGTFDISILDIDNGIFEVKS
TNGDTHLGGEDFDIYLLQEIISHFKKETGIDLSNDRMAVQRIREAAEKAKIELSSTLSTE
INLPFITADAAGPKHIRMPFSRVQLENITAPLIDRTVDPVKKALKDARITASDISDVLLV
GGMSRMPKVADTVKKLFGKDASKAVNPDEAVALGAAIQAAVLSGEVTDVLLLDVTPLSLG
IETLGGVFTKLIPRNSTIPNKKSQIFSTAASGQTSVEVKVFQGERELVKDNKLIGNFTLA
GIPPAPKGTPQIEVTFDIDANGIINVSAKDLASHKDSSITVAGASGLSDTEIDRMVNEAE
RYKNQDRARRNAIETANKADQLANDTENSIKEFEGKLDKTDSQRLKDQISSLRELVSRSQ
AGDEVNDDDVGTKIDNLRTSSMKLFEQLYKNSDNPETKNGRENK*
[0420] ECM10 is encoded by a non-essential gene comprising an ORF
that is 1.935 kbp in size and is located on chromosome V. A
published nucleotide coding sequence of ECM10 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00030
ATGTTACCATCATGGAAAGCCTTTAAAGCACATAATATACTTCGTATTCTGACCCGTTTC
CAGTCAACCAAAATTCCAGATGCAGTTATCGGTATTGATTTAGGTACTACCAATTCTGCG
GTAGCTATTATGGAAGGTAAAGTTCCGAGAATTATCGAAAATGCAGAAGGCTCAAGAACT
ACTCCGTCTGTAGTGGCTTTCACTAAAGACGGAGAACGTTTAGTTGGTGAGCCAGCCAAA
CGACAATCCGTCATAAACTCAGAAAACACTTTGTTTGCTACTAAGCGTTTAATCGGCCGC
CGTTTCGAGGACGCTGAAGTCCAAAGAGATATTAATCAGGTTCCTTTCAAAATCGTCAAG
CATTCTAATGGAGATGCCTGGGTAGAGGCTAGAAACAGAACGTACTCCCCCGCCCAAATA
GGAGGTTTTATCTTAAATAAAATGAAGGAAACAGCGGAGGCTTACTTAGCGAAGAGCGTC
AAAAATGCTGTTGTCACCGTTCCTGCTTACTTCAATGATGCCCAAAGACAAGCTACTAAA
GACGCAGGACAAATTATTGGGCTTAATGTATTACGTGTTGTCAACGAACCAACAGCTGCT
GCCCTAGCTTACGGTCTAGATAAATCAGAGCCAAAAGTCATTGCTGTTTTCGACTTGGGC
GGTGGTACTTTCGATATTTCAATCCTGGACATCGATAACGGTATCTTTGAGGTTAAATCT
ACCAATGGTGACACCCATTTGGGTGGCGAAGATTTTGACATTTATTTGTTGCAAGAAATT
ATTTCTCATTTCAAGAAAGAAACCGGTATCGATTTGAGTAATGACCGTATGGCTGTCCAA
AGAATAAGAGAAGCCGCTGAAAAGGCTAAAATCGAACTGTCTTCTACACTCTCTACAGAA
ATAAACTTGCCTTTCATAACTGCTGATGCTGCAGGCCCAAAGCATATTCGTATGCCCTTT
TCTAGGGTTCAGCTTGAGAATATAACCGCCCCATTGATTGATAGAACGGTTGATCCTGTC
AAAAAAGCACTGAAAGACGCAAGAATTACCGCCTCAGATATATCGGATGTTTTATTAGTT
GGTGGTATGTCAAGGATGCCCAAGGTTGCAGATACTGTAAAGAAATTATTCGGTAAGGAT
GCATCAAAAGCTGTTAACCCTGATGAAGCAGTCGCTTTAGGGGCCGCTATACAGGCTGCG
GTCTTGTCTGGTGAAGTTACCGATGTTTTGTTGCTAGATGTCACTCCCCTATCATTGGGT
ATTGAAACTTTAGGAGGAGTTTTTACAAAATTAATCCCAAGAAATTCTACAATTCCCAAT
AAGAAATCTCAAATTTTTTCAACTGCGGCATCAGGTCAAACATCGGTGGAAGTTAAAGTT
TTCCAAGGTGAGAGGGAGTTAGTCAAGGATAACAAATTAATAGGTAATTTTACTCTTGCG
GGCATTCCTCCAGCTCCAAAAGGTACCCCACAAATTGAAGTCACTTTTGATATCGATGCG
AACGGCATCATCAACGTTTCAGCAAAAGATCTCGCCAGCCACAAAGACTCTTCCATCACT
GTTGCCGGAGCGTCTGGGCTATCTGATACGGAGATTGATCGAATGGTTAATGAAGCGGAA
AGATATAAAAATCAGGATAGAGCCAGAAGGAATGCCATCGAAACCGCTAACAAAGCTGAC
CAGCTAGCTAATGACACAGAAAATTCCATTAAGGAATTCGAAGGTAAGCTAGATAAAACT
GATTCTCAAAGACTAAAAGATCAAATTTCATCCTTAAGGGAATTGGTTTCTCGGAGTCAA
GCTGGAGATGAGGTTAATGATGACGATGTTGGAACAAAAATTGACAATTTGCGAACTTCA
TCGATGAAACTTTTTGAACAGTTATACAAGAACAGTGACAATCCTGAAACTAAGAACGGG
AGAGAAAATAAATAA
[0421] Further information on ECM10 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000000756.
[0422] It will be appreciated that, by "ECM10", we include
fragments or variants thereof having equivalent ECM10-like
activity.
[0423] MDJ1 is another S. cerevisiae helper protein of interest for
the present invention. Mdj1p is a protein involved in folding of
mitochondrially synthesised proteins in the mitochondrial matrix.
It localises to the mitochondrial inner membrane and is a member of
the DnaJ family of molecular chaperones. A published protein
sequence for the protein Mdj1p is as follows:
TABLE-US-00031
MAFQQGVLSRCSGVFRHHVGHSRHINNILYRHAIAFASIAPRIPKSSFHTSAIRNNEAFK
DPYDTLGLKKSATGAEIKKAYYKLAKKYHPDINKEPDAEKKFHDLQNAYEILSDETKRQQ
YDQFGPAAFGGGGAAGGAGGGSGSPFGSQFHDFSGFTSAGGSPFGGINFEDLFGAAFGGG
GRGSGGASRSSSMFRQYRGDPIEIVHKVSFKDAVFGSKNVQLRFSALDPCSTCSGTGMKP
NTHKVSCSTCHGTGTTVHIRGGFQMMSTCPTCNGEGTMKRPQDNCTKCHGEGVQVNRAKT
ITVDLPHGLQDGDVVRIPGQGSYPDIAVEADLKDSVKLSRGDILVRIRVDKDPNFSIKNK
YDIWYDKEIPITTAALGGTVTIPTVEGQKIRIKVAPGTQYNQVISIPNMGVPKTSTIRGD
MKVQYKIVVKKPQSLAEKCLWEALADVTNDDMAKKTMQPGTAAGTAINEEILKKQKQEEE
KHAKKDDDNTLKRLENFITNTFRKIKGDKKN*
[0424] MDJ1 is encoded by a non-essential gene comprising an ORF
that is 1.536 kbp in size and is located on chromosome VI. A
published nucleotide coding sequence of MDJ1 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00032
ATGGCTTTCCAACAAGGTGTATTGTCAAGGTGTTCCGGTGTCTTTAGACACCATGTGGGA
CATTCTCGCCATATCAATAATATTCTTTATAGACATGCCATCGCGTTTGCATCCATCGCT
CCACGAATACCAAAATCTAGCTTCCATACTTCTGCAATCAGAAACAACGAAGCATTCAAG
GACCCGTACGATACTTTAGGCTTGAAGAAATCTGCTACAGGTGCGGAAATCAAAAAAGCA
TACTACAAACTGGCAAAGAAGTACCACCCGGATATCAACAAGGAACCGGATGCTGAGAAG
AAATTCCACGATTTACAGAACGCTTATGAAATTCTGTCAGACGAAACGAAGAGGCAGCAG
TACGATCAATTTGGGCCCGCTGCCTTCGGCGGCGGCGGTGCCGCTGGAGGTGCCGGTGGT
GGTAGTGGCTCTCCCTTTGGTTCCCAATTTCATGATTTCTCAGGATTCACCAGTGCAGGC
GGCTCGCCATTTGGCGGTATCAATTTTGAAGACCTGTTTGGTGCTGCATTTGGTGGTGGT
GGCCGCGGTAGCGGTGGCGCAAGCAGGTCGTCATCTATGTTCAGACAATATAGGGGCGAC
CCAATCGAGATTGTCCATAAAGTGTCTTTCAAGGACGCAGTGTTTGGGTCCAAGAACGTT
CAGTTAAGATTCTCTGCGCTGGACCCTTGTAGTACCTGTTCAGGGACGGGAATGAAACCA
AACACGCATAAGGTCAGTTGTAGCACTTGTCACGGAACAGGAACCACTGTTCACATTAGG
GGCGGATTTCAGATGATGTCGACTTGTCCTACTTGCAACGGTGAAGGTACCATGAAACGG
CCTCAGGACAATTGTACCAAGTGCCATGGTGAGGGTGTTCAGGTCAACAGGGCAAAGACA
ATTACGGTGGACTTGCCACATGGATTACAGGACGGCGACGTGGTCAGGATCCCTGGCCAA
GGCTCATACCCTGACATCGCTGTAGAGGCGGACTTGAAAGATTCAGTCAAGTTATCAAGA
GGTGATATTTTGGTGAGAATTCGTGTCGACAAGGATCCCAACTTTTCGATAAAGAACAAG
TACGATATTTGGTACGACAAGGAGATTCCTATAACCACAGCTGCACTTGGTGGTACTGTC
ACTATCCCCACTGTGGAGGGACAAAAGATCAGGATAAAGGTCGCTCCAGGGACTCAATAC
AATCAAGTGATATCCATTCCTAACATGGGTGTTCCTAAAACATCAACCATTCGCGGTGAT
ATGAAAGTCCAGTACAAGATCGTTGTTAAGAAACCGCAATCGCTGGCAGAAAAATGCTTG
TGGGAGGCACTGGCAGATGTCACCAACGATGACATGGCCAAGAAAACCATGCAACCGGGC
ACAGCCGCGGGTACAGCCATTAATGAAGAGATACTGAAGAAACAAAAACAAGAAGAGGAA
AAACACGCAAAAAAGGATGACGACAACACTTTGAAGAGACTAGAAAATTTCATTACCAAC
ACATTCAGGAAGATCAAAGGTGACAAAAAAAATTAA
[0425] Further information on MDJ1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000001878.
[0426] It will be appreciated that, by "MDJ1", we include fragments
or variants thereof having equivalent MDJ1-like activity.
[0427] MDJ2 is another S. cerevisiae helper protein of interest for
the present invention. Mdj2p is a protein of the mitochondrial
inner membrane. Its function partially overlaps that of Mdj1p,
which is a chaperone involved in folding of mitochondrially
synthesised proteins in the mitochondrial matrix. It is a member of
the DnaJ family. A published protein sequence for the protein Mdj2p
is as follows:
TABLE-US-00033 MVLPIIIGLGVTMVALSVKSGLNAWTVYKTLSPLTIAKLNNIRIENPTA
GYRDALKFKSSLIDEELKNRLNQYQGGFAPRMTEPEALLILDISAREIN
HLDEKLLKKKHRKAMVRNHPDRGGSPYMAAKINEAKEVLERSVLLRKR*
[0428] MDJ2 is encoded by a non-essential gene comprising an ORF
that is 0.441 kbp in size and is located on chromosome XIV. A
published nucleotide coding sequence of MDJ2 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00034
ATGGTTTTGCCTATAATAATTGGTTTGGGCGTGACAATGGTTGCTCTAAGTGTCAAGTCT
GGTCTCAATGCATGGACCGTCTACAAGACCCTGTCCCCTTTAACTATTGCAAAACTAAAT
AACATTCGCATAGAAAACCCGACGGCGGGCTACCGCGATGCACTTAAGTTCAAAAGCTCA
CTGATAGACGAAGAACTGAAAAATAGATTAAACCAGTACCAGGGAGGCTTTGCACCGCGA
ATGACAGAGCCCGAAGCCTTGCTCATCTTGGATATCTCCGCCAGAGAGATTAATCACTTG
GATGAAAAATTACTGAAAAAAAAGCACAGGAAGGCTATGGTTCGTAACCACCCAGACAGA
GGAGGGAGTCCCTACATGGCGGCCAAGATAAATGAGGCGAAAGAAGTTCTCGAAAGAAGT
GTTTTACTAAGAAAGAGATAA
[0429] Further information on MDJ2 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000005272.
[0430] It will be appreciated that, by "MDJ2", we include fragments
or variants thereof having equivalent MDJ2-like activity.
[0431] ERO1 is another S. cerevisiae helper protein of interest for
the present invention. EroIp is a glycoprotein required for
oxidative protein folding in the endoplasmic reticulum. A published
protein sequence for the protein Ero1p is as follows:
TABLE-US-00035
MRLRTAIATLCLTAFTSATSNNSYIATDQTQNAFNDTHFCKVDRNDHVSPSCNVTFNELN
AINENIRDDLSALLKSDFFKYFRLDLYKQCSFWDANDGLCLNRACSVDVVEDWDTLPEYW
QPEILGSFNNDTMKEADDSDDECKFLDQLCQTSKKPVDIEDTINYCDVNDFNGKNAVLID
LTANPERFTGYGGKQAGQIWSTIYQDNCFTIGETGESLAKDAFYRLVSGFHASIGTHLSK
EYLNTKTGKWEPNLDLFMARIGNFPDRVTNMYFNYAVVAKALWKIQPYLPEFSFCDLVNK
EIKNKMDNVISQLDTKIFNEDLVFANDLSLTLKDEFRSRFKNVTKIMDCVQCDRCRLWGK
IQTTGYATALKILFEINDADEFTKQHIVGKLTKYELIALLQTFGRLSESIESVNMFEKMY
GKRLNGSENRLSSFFQNNFFNILKEAGKSIRYTIENINSTKEGKKKTNNSQSHVFDDLKM
PKAEIVPRPSNGTVNKWKKAWNTEVNNVLEAFRFIYRSYLDLPRNIWELSLMKVYKFWNK
FIGVADYVSEETREPISYKLDIQ*
[0432] ERO1 is encoded by an essential gene comprising an ORF that
is 1.692 kbp in size and is located on chromosome XIII. A published
nucleotide coding sequence of ERO1 is as follows, although it will
be appreciated that the sequence can be modified by degenerate
substitutions to obtain alternative nucleotide sequences which
encode an identical protein product:
TABLE-US-00036
ATGAGATTAAGAACCGCCATTGCCACACTGTGCCTCACGGCTTTTACATCTGCAACTTCA
AACAATAGCTACATCGCCACCGACCAAACACAAAATGCCTTTAATGACACTCACTTTTGT
AAGGTCGACAGGAATGATCACGTTAGTCCCAGTTGTAACGTAACATTCAATGAATTAAAT
GCCATAAATGAAAACATTAGAGATGATCTTTCGGCGTTATTAAAATCTGATTTCTTCAAA
TACTTTCGGCTGGATTTATACAAGCAATGTTCATTTTGGGACGCCAACGATGGTCTGTGC
TTAAACCGCGCTTGCTCTGTTGATGTCGTAGAGGACTGGGATACACTGCCTGAGTACTGG
CAGCCTGAGATCTTGGGTAGTTTCAATAATGATACAATGAAGGAAGCGGATGATAGCGAT
GACGAATGTAAGTTCTTAGATCAACTATGTCAAACCAGTAAAAAACCTGTAGATATCGAA
GACACCATCAACTACTGTGATGTAAATGACTTTAACGGTAAAAACGCCGTTCTGATTGAT
TTAACAGCAAATCCGGAACGATTTACAGGTTATGGTGGTAAGCAAGCTGGTCAAATTTGG
TCTACTATCTACCAAGACAACTGTTTTACAATTGGCGAAACTGGTGAATCATTGGCCAAA
GATGCATTTTATAGACTTGTATCCGGTTTCCATGCCTCTATCGGTACTCACTTATCAAAG
GAATATTTGAACACGAAAACTGGTAAATGGGAGCCCAATCTGGATTTGTTTATGGCAAGA
ATCGGGAACTTTCCTGATAGAGTGACAAACATGTATTTCAATTATGCTGTTGTAGCTAAG
GCTCTCTGGAAAATTCAACCATATTTACCAGAATTTTCATTCTGTGATCTAGTCAATAAA
GAAATCAAAAACAAAATGGATAACGTTATTTCCCAGCTGGACACAAAAATTTTTAACGAA
GACTTAGTTTTTGCCAACGACCTAAGTTTGACTTTGAAGGACGAATTCAGATCTCGCTTC
AAGAATGTCACGAAGATTATGGATTGTGTGCAATGTGATAGATGTAGATTGTGGGGCAAA
ATTCAAACTACCGGTTACGCAACTGCCTTGAAAATTTTGTTTGAAATCAACGACGCTGAT
GAATTCACCAAACAACATATTGTTGGTAAGTTAACCAAATATGAGTTGATTGCACTATTA
CAGACTTTCGGTAGATTATCTGAATCTATTGAATCTGTTAACATGTTCGAAAAAATGTAC
GGGAAAAGGTTAAACGGTTCTGAAAACAGGTTAAGCTCATTCTTCCAAAATAACTTCTTC
AACATTTTGAAGGAGGCAGGCAAATCGATTCGTTACACCATAGAGAACATCAATTCCACT
AAAGAAGGAAAGAAAAAGACTAACAATTCTCAATCACATGTATTTGATGATTTAAAAATG
CCCAAAGCAGAAATAGTTCCAAGGCCCTCTAACGGTACAGTAAATAAATGGAAGAAAGCT
TGGAATACTGAAGTTAACAACGTTTTAGAAGCATTCAGATTTATTTATAGAAGCTATTTG
GATTTACCCAGGAACATCTGGGAATTATCTTTGATGAAGGTATACAAATTTTGGAATAAA
TTCATCGGTGTTGCTGATTACGTTAGTGAGGAGACACGAGAGCCTATTTCCTATAAGCTA
GATATACAATAA
[0433] Further information on ERO1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000004599.
[0434] It will be appreciated that, by "ERO1", we include fragments
or variants thereof having equivalent ERO1-like activity.
[0435] ERV2 is another S. cerevisiae helper protein of interest for
the present invention. Erv2p is a flavin-linked sulfhydryl oxidase
localised to the endoplasmic reticulum lumen, involved in
disulphide bond formation within the ER. A published protein
sequence for the protein Erv2p is as follows:
TABLE-US-00037 MKQIVKRSHAIRIVAALGIIGLWMFFSSNELSIATPGLIKAKSGIDEVQG
AAAEKNDARLKEIEKQTIMPLMGDDKVKKEVGRASWKYFHTLLARFPDEP
TPEEREKLHTFIGLYAELYPCGECSYHFVKLIEKYPVQTSSRTAAMWA
GCHIHNKVNEYLKKDIYDCATILEDYDCGCSDSDGKRVSLEKEAKQHG*
[0436] ERV2 is encoded by a non-essential gene comprising an ORF
that is 0.591 kbp in size, located on chromosome XVI. A published
nucleotide coding sequence of ERV2 is as follows, although it will
be appreciated that the sequence can be modified by degenerate
substitutions to obtain alternative nucleotide sequences which
encode an identical protein product:
TABLE-US-00038
ATGAAACAGATAGTCAAAAGAAGCCATGCCATCAGAATAGTTGCAGCATTAGGAATCATA
GGCCTGTGGATGTTTTTCTCGTCTAATGAACTATCCATCGCTACGCCGGGCCTAATCAAG
GCGAAGTCTGGTATAGATGAAGTGCAAGGGGCGGCTGCTGAGAAGAACGACGCTCGGTTG
AAAGAGATCGAGAAGCAAACCATTATGCCATTGATGGGCGATGACAAGGTGAAGAAGGAA
GTGGGCAGGGCGTCGTGGAAGTACTTCCATACCCTGCTGGCCCGTTTTCCGGACGAGCCT
ACTCCTGAAGAAAGAGAGAAACTGCACACGTTTATTGGGTTGTATGCAGAACTCTATCCA
TGCGGGGAATGTTCATATCACTTTGTAAAGTTGATTGAGAAGTATCCCGTACAGACATCT
AGCAGGACGGCTGCCGCAATGTGGGGATGCCACATTCACAACAAGGTGAACGAATACCTA
AAGAAAGACATATATGACTGTGCTACCATCCTGGAGGACTACGATTGTGGATGTAGTGAC
AGCGACGGTAAACGCGTGTCTCTCGAGAAGGAGGCTAAACAGCACGGTTGA
[0437] Further information on ERV2 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000006241.
[0438] It will be appreciated that, by "ERV2", we include fragments
or variants thereof having equivalent ERV2-like activity.
[0439] EUG1 is another S. cerevisiae helper protein of interest for
the present invention. Eug1p is a protein disulphide isomerase of
the endoplasmic reticulum lumen, with an overlapping function with
Pdi1p. It may interact with nascent polypeptides in the ER. A
published protein sequence for the protein Eug1p is as follows:
TABLE-US-00039
MQVTTRFISAIVSFCLFASFTLAENSARATPGSDLLVLTEKKFKSFIESHPLVLVEFFAP
WCLHSQILRPHLEEAASILKEHNVPVVQIDCEANSMVCLQQTINTYPTLKIFKNGRIFDG
QVYRGVKITDEITQYMIQLYEASVIYLNSEDEIQPYLENATLPVVINRGLTGLNETYQEV
ALDLAEDYVFLSLLDSEDKSLSIHLPNTTEPILFDGNVDSLVGNSVALTQWLKVVILPYF
TDIEPDLFPKYISSNLPLAYFFYTSEEELEDYTDLFTQLGKENRGQINFIALNSTMFPHH
VRFLNMREQFPLFAIHNMINNLKYGLPQLPEEEYAKLEKPQPLDRDMIVQLVKDYREGTA
KPIVKSEEIPKEQKSNVYKIVGKTHDDIVHDDDKDVLVKYYATWCIHSKRFAPIYEEIAN
VLASDESVRDKILIAEVDSGANDILSFPVTGYPTIALYPAGNNSKPIIFNKIRNLEDVFE
FIKESGTHHIDGQAIYDKLHQAKDSEVSTEDTVHDEL*
[0440] EUG1 is encoded by a non-essential gene comprising an ORF
that is 1.554 kbp in size and is located on chromosome IV. A
published nucleotide coding sequence of EUG1 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00040
ATGCAAGTGACCACAAGATTTATATCTGCGATAGTCTCGTTTTGCCTGTTTGCTTCTTTC
ACGTTGGCTGAAAACAGCGCAAGAGCTACGCCGGGATCAGATTTACTCGTTCTAACAGAG
AAGAAATTTAAATCATTCATCGAATCTCATCCGTTAGTCCTCGTCGAGTTTTTTGCTCCA
TGGTGTTTGCATTCTCAGATCTTACGCCCTCACTTAGAAGAGGCCGCCTCTATTTTAAAG
GAGCATAACGTCCCAGTTGTTCAAATTGATTGTGAGGCTAACAGTATGGTTTGCCTGCAA
CAAACTATAAATACCTACCCAACCTTGAAAATCTTTAAAAATGGTCGTATTTTTGATGGT
CAAGTCTATCGCGGTGTCAAGATCACCGATGAAATCACTCAGTACATGATTCAGCTATAC
GAGGCTTCTGTCATTTATTTAAATTCCGAAGATGAAATCCAACCATACTTGGAAAATGCA
ACTTTACCAGTAGTAATAAACAGAGGCTTGACAGGCTTGAATGAAACGTATCAAGAAGTC
GCACTGGACCTTGCTGAGGATTACGTCTTTTTATCCCTTCTAGATTCAGAAGATAAGTCA
TTATCAATCCACTTGCCAAACACTACAGAACCAATTCTGTTTGATGGAAATGTAGACTCT
TTGGTCGGAAATTCCGTTGCTCTAACTCAGTGGTTAAAAGTGGTAATTTTACCTTACTTT
ACCGACATCGAACCTGATCTCTTCCCCAAGTACATTTCTAGCAATTTGCCGTTGGCTTAC
TTCTTTTATACTTCTGAGGAAGAATTGGAAGATTACACTGATCTTTTCACGCAGTTAGGT
AAGGAAAATCGTGGCCAAATAAATTTCATTGCATTAAACTCTACAATGTTCCCACACCAC
GTTAGATTCCTAAATATGAGAGAACAGTTCCCATTATTTGCTATCCATAATATGATCAAT
AATCTGAAATATGGTTTACCACAACTACCAGAAGAAGAGTACGCGAAATTAGAAAAACCA
CAACCACTAGACAGAGATATGATCGTTCAGTTGGTAAAAGATTACCGTGAAGGTACTGCC
AAGCCAATTGTTAAGTCAGAAGAGATTCCAAAAGAACAAAAGTCCAATGTTTATAAAATA
GTTGGGAAGACACATGACGACATTGTTCATGATGATGACAAGGATGTCCTTGTCAAATAT
TACGCGACATGGTGTATTCATAGTAAAAGGTTTGCGCCTATTTACGAAGAAATTGCAAAT
GTCTTAGCATCTGATGAATCTGTTCGCGATAAAATCTTGATCGCCGAAGTAGATTCAGGG
GCAAATGATATCTTAAGTTTTCCTGTGACAGGATATCCAACCATTGCTTTGTATCCTGCC
GGAAATAACTCTAAGCCTATTATCTTCAATAAAATTAGAAATTTGGAAGATGTTTTCGAA
TTTATCAAGGAATCAGGTACACATCACATTGACGGCCAGGCAATTTATGATAAATTGCAC
CAGGCCAAGGATTCTGAAGTGTCTACTGAAGATACCGTACATGATGAATTATAA
[0441] Further information on EUG1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000002926.
[0442] It will be appreciated that, by "EUG1", we include fragments
or variants thereof having equivalent EUG1-like activity.
[0443] MPD1 is another S. cerevisiae helper protein of interest for
the present invention. Mpd1p is a member of the protein disulphide
isomerase (PDI) family. Its over-expression suppresses the defect
in maturation of carboxypeptidase Y, and defects in other essential
Pdi1p functions that can be caused by PDI1 deletion. A published
protein sequence for the protein Mpd1p is as follows:
TABLE-US-00041 MLFLNIIKLLLGLFIMNEVKAQNFYDSDPHISELTPKSFDKAIHNTNYTS
LVEFYAPWCGHCKKLSSTFRKAAKRLDGVVQVAAVNCDLNKNKALCAKYD
VNGFPTLMVFRPPKIDLSKPIDNAKKSFSAHANEVYSGARTLAPIVDFSL
SRIRSYVKKFVRIDTLGSLLRKSPKLSVVLFSKQDKISPVYKSIALDWLG
KFDFYSISNKKLKQLTDMNPTYEKTPEIFKYLQKVIPEQRQSDKSKLVVF
DADKDKFWEYEGNSINKNDISKFLRDTFSITPNEGPFSRRSEYIAYLKTG
KKPIKKNHSSSGNKHDEL*
[0444] MPD1 is encoded by a non-essential gene comprising an ORF
that is 0.957 kbp in size and is located on chromosome XV. A
published nucleotide coding sequence of MPD1 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00042
ATGTTATTTCTTAATATTATTAAGCTCCTTTTGGGACTTTTTATTATGAATGAAGTAAAG
GCGCAAAACTTTTACGATTCCGATCCTCATATATCAGAGTTAACGCCAAAAAGCTTCGAT
AAAGCGATCCATAACACAAATTACACATCATTAGTGGAATTTTATGCTCCGTGGTGCGGC
CATTGTAAGAAGCTCTCTAGTACGTTCCGCAAGGCAGCAAAAAGATTGGATGGTGTAGTC
CAAGTTGCTGCTGTAAACTGTGACCTTAACAAGAATAAGGCTTTGTGTGCTAAATACGAC
GTAAACGGATTTCCCACGTTAATGGTATTTAGGCCCCCAAAAATTGACCTATCTAAGCCA
ATAGATAACGCCAAAAAAAGTTTCAGCGCTCATGCCAATGAAGTGTACTCAGGTGCAAGA
ACTCTCGCGCCTATTGTTGATTTTTCTCTTTCAAGAATAAGGTCATATGTCAAAAAGTTT
GTCCGTATAGATACACTTGGCTCTTTACTTAGAAAGTCACCCAAACTTTCCGTGGTGTTG
TTTTCCAAACAAGACAAAATTTCACCGGTTTATAAAAGCATTGCCCTTGATTGGTTAGGA
AAGTTCGATTTTTATTCAATTTCAAACAAAAAACTCAAGCAACTAACCGATATGAACCCA
ACATATGAAAAAACTCCTGAGATTTTCAAATATTTGCAGAAGGTCATTCCTGAACAGCGA
CAGAGCGATAAAAGTAAGCTTGTCGTTTTTGATGCAGACAAAGATAAATTTTGGGAGTAT
GAAGGGAACTCAATCAACAAAAATGACATTTCCAAATTTCTGCGGGACACTTTTAGTATT
ACCCCCAATGAGGGTCCTTTTAGTAGACGTTCTGAATATATTGCTTACTTAAAAACTGGC
AAGAAGCCAATTAAAAAGAACCATTCCTCCTCAGGAAACAAGCACGACGAATTGTAG
[0445] Further information on MPD1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000005814.
[0446] It will be appreciated that, by "MPD1", we include fragments
or variants thereof having equivalent MPD1-like activity.
[0447] MPD2 is another S. cerevisiae helper protein of interest for
the present invention. Mpd2p is a member of the protein disulphide
isomerase (PDI) family. It exhibits chaperone activity. Its
overexpression suppresses the lethality of a PDI1 deletion but does
not complement all Pdi1p functions. It undergoes oxidation by
Ero1p. A published protein sequence for the protein Mpd2p is as
follows:
TABLE-US-00043 MKLHGFLFSVLSTCVVILPALAYSEAVTMVKSIEQYFDICNRNDSYTMIK
YYTSWCQHCKTLAPVYEELGELYAKKANKDDTPINFLEVNCEFFGPTLCT
DLPGFPIIELVKPRTKPLVLPKLDWSSMKFHERLWQRIKTWFNNPKYQLD
TSRVVRFEGSRNLKSLSNFIDTVRSKDTEERFIEHIFDDSRNCNEELRSQ
QLLCKAGKEYYSDTLSKLYGDVNGLEKERRRLEALIKQNGDDLSKEVKEK
LKIIRLQLSLLSHIEDQLEDTSSHDEL*
[0448] MPD2 is encoded by a non-essential gene comprising an ORF
that is 0.834 kbp in size and is located on chromosome XV. A
published nucleotide coding sequence of MPD2 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00044
ATGAAATTGCACGGCTTTTTATTTTCCGTATTATCAACATGCGTCGTCATTTTACCAGCG
TTGGCCTACAGTGAAGCTGTCACGATGGTCAAGTCGATTGAGCAGTACTTCGATATCTGC
AATAGGAATGATTCTTACACAATGATAAAATACTACACTTCTTGGTGCCAACATTGTAAA
ACTCTGGCCCCAGTATACGAAGAGCTTGGTGAGCTATACGCCAAAAAAGCTAATAAAGAT
GATACCCCAATTAACTTCCTTGAAGTTAACTGTGAATTCTTCGGGCCAACTTTATGTACC
GACTTGCCTGGATTTCCAATAATTGAACTGGTCAAACCTCGTACTAAGCCCTTAGTTCTT
CCGAAGCTCGATTGGTCGTCTATGAAATTTCATGAAAGACTATGGCAAAGAATCAAGACG
TGGTTCAACAATCCTAAGTACCAACTGGATACGTCTAGGGTTGTTCGTTTTGAAGGGAGT
AGGAACCTAAAGAGTTTAAGCAACTTTATCGATACTGTAAGAAGTAAAGATACAGAAGAA
AGATTCATAGAACATATTTTCGATGATTCTAGGAATTGCAATGAAGAATTACGTTCTCAA
CAGCTTCTGTGTAAAGCTGGTAAAGAATACTACTCTGATACTTTATCTAAATTATACGGT
GACGTGAATGGGCTGGAAAAGGAAAGGCGAAGACTAGAAGCTTTAATTAAGCAAAATGGA
GATGACTTGAGTAAAGAAGTTAAAGAAAAACTGAAAATCATTCGTCTACAATTGAGCCTA
TTATCACACATAGAAGACCAGTTAGAAGATACCAGTAGTCATGACGAGCTTTGA
[0449] Further information on MPD2 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000005448.
[0450] It will be appreciated that, by "MPD2", we include fragments
or variants thereof having equivalent MPD2-like activity.
[0451] EPS1 is another S. cerevisiae helper protein of interest for
the present invention. Eps1p is a Pdi1p (protein disulphide
isomerase)-related protein involved in endoplasmic reticulum
retention of resident ER proteins. A published protein sequence for
the protein Eps1p is as follows:
TABLE-US-00045
MKMNLKRLVVTFFSCITFLLKFTIAAAEPPEGFPEPLNPTNFKEELSKGLHIIDFYSPYC
PHCKHLAPVWMETWEEFKEESKTLNITFSQVNCIESADLCGDENIEYFPEIRLYNPSGYI
KSFTETPRTKESLIAFARRESMDPNNLDTDLDSAKSESQYLEGFDFLELIAGKATRPHLV
SFWPTKDMKNSDDSLEFKNCDKCHEFQRTWKIISRQLAVDDINTGHVNCESNPTICEELG
FGDLVKITNHRADREPKVALVLPNKTSNNLFDYPNGYSAKSDGYVDFARRTFTNSKFPNI
TEGELEKKANRDIDFLQERGRVTNNDIHLVFSYDPETVVIEDFDILEYLIEPLSKIPNIY
LHQIDKNLINLSRNLFGRMYEKINYDASQTQKVFNKEYFTMNTVTQLPTFFMFKDGDPIS
YVFPGYSTTEMRNIDAIMDWVKKYSNPLVTEVDSSNLKKLISFQTKSYSDLAIQLISSTD
HKHIKGSNKLIKNLLLASWEYEHIRMENNFEEINERRARKADGIKKIKEKKAPANKIVDK
MREEIPHMDQKKLLLGYLDISKEKNFFRKYGITGEYKIGDVIIIDKSNNYYYNKDNFGNS
LTSNNPQLLREAFVSLNIPSKALYSSKLKGRLINSPFHNVLSFLDIIHGNGMPGYLIVIV
LFIAILKGPSIYRRYKVRKHYRAKRNAVGILGNMEKKKNQD*
[0452] EPS1 is a non-essential gene comprising an ORF that is 2.106
kbp in size and is located on chromosome IX. A published nucleotide
coding sequence of EPS1 is as follows, although it will be
appreciated that the sequence can be modified by degenerate
substitutions to obtain alternative nucleotide sequences which
encode an identical protein product:
TABLE-US-00046
ATGAAAATGAATCTGAAAAGGCTCGTAGTTACCTTCTTCTCATGCATCACCTTTCTGCTG
AAATTCACTATAGCCGCCGCTGAACCACCAGAGGGCTTTCCAGAGCCCTTAAATCCAACA
AACTTCAAAGAAGAGCTATCTAAGGGGCTGCATATTATTGACTTCTATAGTCCATACTGT
CCGCACTGCAAACATTTAGCACCTGTTTGGATGGAAACATGGGAGGAGTTTAAAGAGGAG
AGCAAAACACTGAACATAACATTTTCACAGGTTAACTGCATCGAGAGCGCCGATTTGTGT
GGAGATGAAAATATTGAATACTTCCCTGAAATTAGACTTTATAACCCCTCAGGATACATC
AAATCGTTCACTGAAACACCGAGGACCAAAGAATCATTAATTGCATTTGCACGCAGGGAG
TCTATGGACCCAAATAACCTCGATACTGATCTGGATTCTGCTAAAAGTGAGAGCCAGTAT
CTCGAAGGCTTTGATTTTCTCGAGCTGATCGCTGGTAAGGCGACTAGGCCACATTTGGTT
TCCTTCTGGCCAACAAAAGATATGAAAAATAGCGATGATTCACTAGAATTCAAAAACTGT
GACAAATGCCATGAATTCCAAAGGACTTGGAAGATCATTTCAAGACAGTTAGCCGTGGAT
GATATCAACACGGGCCACGTTAATTGCGAATCTAATCCAACAATCTGTGAAGAACTGGGC
TTTGGCGACTTGGTGAAAATAACCAACCACAGAGCCGATAGAGAACCCAAGGTAGCATTA
GTCCTACCCAATAAAACCTCAAATAATTTGTTCGACTATCCCAATGGCTACTCAGCGAAG
TCAGATGGCTATGTAGATTTTGCCAGGAGGACTTTTACAAACAGTAAATTTCCCAATATA
ACAGAAGGGGAGCTCGAAAAAAAAGCAAACAGAGACATTGATTTTCTGCAAGAAAGGGGA
CGAGTAACTAATAATGATATCCATTTAGTTTTTTCATATGACCCCGAAACTGTTGTTATT
GAAGATTTTGACATTTTGGAGTATTTAATCGAGCCTTTGTCAAAAATTCCAAACATATAT
TTGCACCAAATTGACAAGAATCTAATAAATTTGTCACGTAATCTTTTTGGAAGAATGTAT
GAAAAGATCAACTACGACGCCAGCCAAACTCAAAAGGTTTTTAACAAAGAATACTTTACT
ATGAATACGGTTACGCAACTCCCAACTTTTTTCATGTTTAAAGATGGTGATCCCATATCC
TATGTTTTCCCCGGATACTCCACAACAGAAATGAGAAATATTGATGCCATTATGGATTGG
GTAAAAAAGTATTCTAATCCCTTAGTTACCGAAGTTGACTCTTCTAATTTGAAAAAATTA
ATTTCCTTCCAAACCAAGAGCTACAGTGATTTAGCAATTCAGTTAATAAGTAGCACTGAC
CACAAACATATCAAAGGAAGCAACAAGCTTATTAAAAACTTGCTCCTCGCAAGTTGGGAG
TATGAACATATTCGGATGGAAAATAACTTCGAAGAAATTAATGAGAGAAGGGCAAGGAAA
GCAGACGGGATCAAGAAAATAAAGGAAAAAAAGGCTCCGGCTAACAAAATTGTTGATAAA
ATGCGTGAAGAGATTCCCCATATGGATCAAAAAAAATTGTTATTAGGATATTTAGATATT
TCAAAGGAGAAGAATTTTTTTAGAAAATATGGTATTACTGGAGAATATAAAATTGGTGAT
GTGATTATCATTGATAAATCAAATAATTACTACTACAATAAAGATAATTTTGGCAACTCC
TTGACTTCTAACAACCCTCAATTGCTGAGAGAAGCATTCGTGTCCTTAAATATTCCATCA
AAAGCTCTATACAGCTCTAAGTTGAAGGGGAGATTGATAAATTCTCCATTCCATAATGTC
CTCAGTTTCCTAGACATAATCCACGGGAACGGCATGCCCGGTTACTTAATTGTTATTGTT
TTGTTTATCGCAATACTCAAAGGTCCATCTATTTACAGAAGATACAAAGTAAGGAAACAC
TATAGGGCGAAAAGGAACGCTGTCGGTATCCTAGGAAATATGGAGAAAAAAAAAAATCAA
GATTAA
[0453] Further information on EPS1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000001267.
[0454] It will be appreciated that, by "EPS1", we include fragments
or variants thereof having equivalent EPS1-like activity.
[0455] PDI, or a fragment or variant thereof having an equivalent
ability to catalyse the formation of disulphide bonds within the
lumen of the endoplasmic reticulum (ER), is another S. cerevisiae
helper protein of interest for the present invention. By "PDI" we
include any protein having the ability to reactivate the
ribonuclease activity against RNA of scrambled ribonuclease as
described in EP 0 746 611 and Hillson et al, 1984, Methods
Enzymol., 107, 281-292.
[0456] PDI is an enzyme which typically catalyses thiol:disulphide
interchange reactions, and is a major resident protein component of
the ER lumen in secretory cells. A body of evidence suggests that
it plays a role in secretory protein biosynthesis (Freedman, 1984,
Trends Biochem. Sci., 9, 438-41) and this is supported by direct
cross-linking studies in situ (Roth and Pierce, 1987, Biochemistry,
26, 4179-82). The finding that microsomal membranes deficient in
PDI show a specific defect in cotranslational protein disulphide
(Bulleid and Freedman, 1988, Nature, 335, 649-51) implies that the
enzyme functions as a catalyst of native disulphide bond formation
during the biosynthesis of secretory and cell surface proteins.
This role is consistent with what is known of the enzyme's
catalytic properties in vitro; it catalyzes thiol: disulphide
interchange reactions leading to net protein disulphide formation,
breakage or isomerisation, and can typically catalyze protein
folding and the formation of native disulphide bonds in a wide
variety of reduced, unfolded protein substrates (Freedman et al.,
1989, Biochem. Soc. Symp., 55, 167-192). PDI also functions as a
chaperone since mutant PDI lacking isomerase activity accelerates
protein folding (Hayano et al, 1995, FEBS Letters, 377, 505-511).
Recently, sulphydryl oxidation, not disulphide isomerisation was
reported to be the principal function of Protein Disulphide
Isomerase in S. cerevisiae (Solovyov et al., 2004, J. Biol. Chem.,
279 (33) 34095-34100). The DNA and amino acid sequence of the
enzyme is known for several species (Scherens et al, 1991, Yeast,
7, 185-193; Farquhar et al, 1991, Gene, 108, 81-89; EP074661;
EP0293793; EP0509841) and there is increasing information on the
mechanism of action of the enzyme purified to homogeneity from
mammalian liver (Creighton et al, 1980, J. Mol. Biol., 142, 43-62;
Freedman et al, 1988, Biochem. Soc. Trans., 16, 96-9; Gilbert,
1989, Biochemistry, 28, 7298-7305; Lundstrom and Holmgren, 1990, J.
Biol. Chem., 265, 9114-9120; Hawkins and Freedman, 1990, Biochem.
J., 275, 335-339). Of the many protein factors currently implicated
as mediators of protein folding, assembly and translocation in the
cell (Rothman, 1989, Cell, 59, 591-601), PDI has a well-defined
catalytic activity.
[0457] The deletion or inactivation of the endogenous PDI gene in a
host results in the production of an inviable host. In other words,
the endogenous PDI gene is an "essential" gene.
[0458] PDI is readily isolated from mammalian tissues and the
homogeneous enzyme is a homodimer (2.times.57 kD) with
characteristically acidic pI (4.0-4.5) (Hinson et al, 1984, op.
cit.). The enzyme has also been purified from wheat and from the
alga Chlamydomonas reinhardii (Kaska et al, 1990, Biochem. J., 268,
63-68), rat (Edman et al, 1985, Nature, 317, 267-270), bovine
(Yamauchi et al, 1987, Biochem. Biophys. Res. Comm., 146,
1485-1492), human (Pihlajaniemi et al, 1987, EMBO J., 6, 643-9),
yeast (Scherens et al, supra; Farquhar et al, op. cit.) and chick
(Parkkonen et al, 1988, Biochem. J., 256, 1005-1011). The proteins
from these vertebrate species show a high degree of sequence
conservation throughout and all show several overall features first
noted in the rat PDI sequence (Edman et al., 1985, op. cit.).
[0459] Preferred PDI sequences include those from humans and those
from yeast species, such as S. cerevisiae.
[0460] A yeast protein disulphide isomerase precursor, PDI1, can be
found as Genbank accession no. CAA42373 or BAA00723. It has the
following sequence of 522 amino acids:
TABLE-US-00047 1 mkfsagavls wsslllassv faqqeavape dsavvklatd
sfneyiqshd lvlaeffapw 61 cghcknmape yvkaaetlve knitlaqidc
tenqdlcmeh nipgfpslki fknsdvnnsi 121 dyegprtaea ivqfmikqsq
pavavvadlp aylanetfvt pvivqsgkid adfnatfysm 181 ankhfndydf
vsaenadddf klsiylpsam depvvyngkk adiadadvfe kwlqvealpy 241
fgeidgsvfa qyvesglplg ylfyndeeel eeykplftel akknrglmnf vsidarkfgr
301 hagnlnmkeq fplfaihdmt edlkyglpql seeafdelsd kivleskaie
slvkdflkgd 361 aspivksqei fenqdssvfq lvgknhdeiv ndpkkdvlvl
yyapwcghck rlaptyqela 421 dtyanatsdv liakldhten dvrgvviegy
ptivlypggk ksesvvyqgs rsldslfdfi 481 kenghfdvdg kalyeeaqek
aaeeadadae ladeedaihd el
[0461] An alternative yeast protein disulphide isomerase sequence
can be found as Genbank accession no. CAA38402. It has the
following sequence of 530 amino acids
TABLE-US-00048 1 mkfsagavls wsslllassv faqqeavape dsavvklatd
sfneyiqshd lvlaeffapw 61 cghcknmape yvkaaetlve knitlaqidc
tenqdlcmeh nipgfpslki fknrdvnnsi 121 dyegprtaea ivqfmikqsq
pavavvadlp aylanetfvt pvivqsgkid adfnatfysm 181 ankhfndydf
vsaenadddf klsiylpsam depvvyngkk adiadadvfe kwlqvealpy 241
fgeidgsvfa qyvesglplg ylfyndeeel eeykplftel akknrglmnf vsidarkfgr
301 hagnlnmkeq fplfaihdmt edlkyglpql seeafdelsd kivleskaie
slvkdflkgd 361 aspivksqei fenqdssvfq lvgknhdeiv ndpkkdvlvl
yyapwcghck rlaptyqela 421 dtyanatsdv liakldhten dvrgvviegy
ptivlypggk ksesvvyqgs rsldslfdfi 481 kenghfdvdg kalyeeaqek
aaeeaeadae aeadadaela deedaihdel
[0462] The following alignment of these sequences (the sequence of
Genbank accession no. CAA42373 or BAA00723 first, the sequence of
Genbank accession no. CAA38402 second) shows that the differences
between these two sequences are a single amino acid difference at
position 114 (highlighted in bold) and that the sequence defined by
Genbank accession no. CAA38402 contains the additional amino acids
EADAEAEA at positions 506-513.
TABLE-US-00049 1 mkfsagavls wsslllassv faqqeavape dsavvklatd
sfneyiqshd lvlaeffapw 1 mkfsagavls wsslllassv faqqeavape dsavvklatd
sfneyiqshd lvlaeffapw 61 cghcknmape yvkaaetlve knitlaqidc
tenqdlcmeh nipgfpslki fknsdvnnsi 61 cghcknmape yvkaaetlve
knitlaqidc tenqdlcmeh nipgfpslki fknrdvnnsi 121 dyegprtaea
ivqfmikqsq pavavvadlp aylanetfvt pvivqsgkid adfnatfysm 181
dyegprtaea ivqfmikqsq pavavvadlp aylanetfvt pvivqsgkid adfnatfysm
181 ankhfndydf vsaenadddf klsiylpsam depvvyngkk adiadadvfe
kwlqvealpy 181 ankhfndydf vsaenadddf klsiylpsam depvvyngkk
adiadadvfe kwlqvealpy 241 fgeidgsvfa qyvesglplg ylfyndeeel
eeykplftel akknrglmnf vsidarkfgr 241 fgeidgsvfa qyvesglplg
ylfyndeeel eeykplftel akknrglmnf vsidarkfgr 301 hagnlnmkeq
fplfaihdmt edlkyglpql seeafdelsd kivleskaie slvkdflkgd 301
hagnlnmkeq fplfaihdmt edlkyglpql seeafdelsd kivleskaie slvkdflkgd
361 aspivksqei fenqdssvfq lvgknhdeiv ndpkkdvlvl yyapwcghck
rlaptyqela 361 aspivksqei fenqdssvfq lvgknhdeiv ndpkkdvlvl
yyapwcghck rlaptyqela 421 dtyanatsdv liakldhten dvrgvviegy
ptivlypggk ksesvvyqgs rsldslfdfi 421 dtyanatsdv liakldhten
dvrgvviegy ptivlypggk ksesvvyqgs rsldslfdfi 481 kenghfdvdg
kalyeeaqek aaeea***** ***dadaela deedaihdel 481 kenghfdvdg
kalyeeaqek aaeeaeadae aeadadaela deedaihdel
[0463] It will be appreciated that, by "PDI" and "PDI1", we include
fragments or variants thereof having equivalent PDI-like activity
and PDI1-like activity, respectively.
[0464] DER1 is another S. cerevisiae helper protein of interest for
the present invention. Der1p is an endoplasmic reticulum membrane
protein, required for the protein degradation process associated
with the ER, and is involved in the retrograde transport of
misfolded or unassembled proteins. A published protein sequence for
the protein Der1p is as follows:
TABLE-US-00050 MDAVILNLLGDIPLVTRLWTIGCLVLSGLTSLRIVDPGKVVYSYDLVF
KKGQYGRLLYSIFDYGAFNWISMINIFVSANHLSTLENSFNLRRKFC
WIIFLLLVILVKMTSIEQPAASLGVLLHENLVYYELKKNGNQMNVRFF
GAIDVSPSIFPIYMNAVMYFVYKRSWLEIAMNFMPGHVIYYMDDIIGKIY
GIDLCKSPYDWFRNTETP*
[0465] DER1 is encoded by a non-essential gene comprising an ORF
that is 0.636 kbp in size and is located on chromosome II. A
published nucleotide coding sequence of DER1 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00051 ATGGATGCTGTAATACTGAATCTCTTAGGCGACATTCCTTTGGTCACA
AGATTATGGACAATTGGCTGTCTTGTACTATCAGGTCTCACAAGTCTCC
GGATTGTGGATCCAGGGAAGGTAGTGTACAGTTATGATTTAGTATTCAA
AAAGGGACAATATGGAAGACTACTTTATTCGATATTCGATTACGGCGCA
TTTAATTGGATATCCATGATAAACATCTTTGTCAGCGCTAATCACTTATC
AACTTTGGAAAACTCATTCAATCTGAGAAGAAAATTCTGTTGGATAATAT
TTTTACTGTTGGTGATACTGGTAAAGATGACCAGCATTGAACAACCTGC
AGCATCACTCGGTGTGTTATTGCATGAGAATCTCGTGTACTACGAACTG
AAAAAGAACGGAAACCAAATGAACGTACGATTCTTCGGTGCCATTGAT
GTTTCACCATCTATATTCCCAATCTACATGAATGCAGTAATGTATTTTGT
ATATAAGCGTAGCTGGTTAGAAATTGCCATGAATTTCATGCCAGGTCAC
GTAATTTACTACATGGATGATATAATAGGGAAGATTTATGGCATCGATTT
GTGTAAATCTCCGTACGACTGGTTCCGCAACACTGAAACACCCTAA
[0466] Further information on DER1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000000405.
[0467] It will be appreciated that, by "DER1", we include fragments
or variants thereof having equivalent DER1-like activity.
[0468] DER3 is another S. cerevisiae helper protein of interest for
the present invention and is also known as HRD1. Der3p is a
ubiquitin-protein ligase required for endoplasmic
reticulum-associated degradation (BRAD) of misfolded proteins. It
is genetically linked to the unfolded protein response (UPR) and is
thought to be regulated through association with Hrd3p. It contains
an H2 ring finger. A published protein sequence for the protein
Der3p is as follows:
TABLE-US-00052 MVPENRRKQLAIFVVVTYLLTFYCVYSATKTSVSFLQVTLKLNEGF
NLMVLSIFILLNSTLLWQLLTKLLFGELRLIEHEHIFERLPFTIINTLFM
SSLFHERYFFTVAFFGLLLLYLKVFHWILKDRLEALLQSINDSTTMK
TLIFSRFSFNLVLLAVVDYQIITRCISSIYTNQKSDIESTSLYLIQVME
FTMLLIDLLNLFLQTCLNFWEFYRSQQSLSNENNHIVHGDPTDENTVE
SDQSQPVLNDDDDDDDDDRQFTGLEGKFMYEKAIDVFTRFLKTAL
HLSMLIPFRMPMMLLKDVVWDILALYQSGTSLWKIWRNNKQLDDTL
VTVTVEQLQNSANDDNICIICMDELIHSPNQQTWKNKNKKPKRLPCG
HILHLSCLKNWMERSQTCPICRLPVFDEKGNVVQTTFTSNSDITTQTT
VTDSTGIATDQQGFANEVDLLPTRTTSPDIRIVPTQNIDTLAMRTRSTS
TPSPTWYTFPLHKTGDNSVGSSRSAYEFLITNSDEKENGIPVKLTIEN
HEVNSLHGDGGEQIAKKIVIPDKFIQHI*
[0469] DER3 is encoded by a non-essential gene comprising an ORF
that is 1.656 kbp in size and is located on chromosome XV. A
published nucleotide coding sequence of DER3 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00053
ATGGTGCCAGAAAATAGAAGGAAACAGTTGGCAATTTTTGTAGTTGTCACATATTTGCTC
ACATTTTATTGCGTGTATTCAGCCACCAAGACAAGCGTTTCCTTTTTGCAAGTAACACTG
AAGCTAAATGAAGGCTTCAATCTAATGGTTTTGTCGATATTCATCTTATTAAATTCTACC
TTACTATGGCAACTCCTAACGAAACTATTATTTGGTGAACTGAGGCTTATTGAGCATGAG
CACATTTTTGAAAGGTTACCATTTACCATTATAAACACCTTGTTTATGTCCTCACTGTTC
CACGAACGGTATTTTTTCACAGTGGCATTTTTTGGACTATTACTACTCTATCTGAAAGTT
TTCCATTGGATTTTAAAGGATAGGCTGGAGGCCTTATTACAGTCAATAAATGATTCCACC
ACAATGAAAACCCTTATCTTTAGTAGATTCTCATTTAACCTCGTACTATTGGCGGTTGTA
GACTACCAGATAATAACACGATGCATCTCCTCCATATATACAAACCAAAAGAGTGATATT
GAATCCACATCCCTTTACCTGATACAAGTAATGGAGTTTACCATGCTTTTGATTGATTTG
CTAAATTTATTCCTACAGACTTGTTTGAATTTCTGGGAATTTTATCGCTCACAACAAAGT
CTGTCTAATGAGAACAACCATATTGTCCATGGCGATCCTACAGATGAAAACACGGTTGAG
TCTGATCAATCTCAGCCAGTGCTGAATGACGACGACGATGACGACGATGATGATAGACAA
TTTACCGGCCTGGAGGGTAAATTCATGTATGAAAAAGCAATTGACGTATTCACAAGATTC
TTAAAAACGGCACTTCATTTGTCTATGCTAATACCATTTAGGATGCCTATGATGCTTTTG
AAAGATGTGGTGTGGGATATCTTGGCACTATATCAAAGTGGCACAAGTTTGTGGAAAATC
TGGAGAAATAACAAACAGCTCGACGACACTCTTGTCACTGTCACCGTAGAACAGCTACAA
AATTCTGCAAATGATGACAATATTTGTATCATTTGTATGGATGAGTTAATACATTCTCCA
AACCAGCAGACGTGGAAGAATAAAAACAAGAAACCCAAAAGGTTACCTTGTGGCCACATA
CTTCATTTGTCGTGTTTAAAGAATTGGATGGAACGTTCTCAGACTTGTCCTATTTGTAGA
TTGCCTGTCTTTGATGAAAAAGGTAATGTTGTGCAAACGACTTTCACTTCCAATAGTGAT
ATCACGACACAGACCACCGTAACAGATAGCACTGGGATAGCGACAGATCAACAAGGTTTC
GCAAACGAAGTAGATCTACTTCCCACAAGAACAACTTCCCCTGATATAAGGATAGTGCCT
ACTCAAAATATAGACACATTAGCAATGAGAACAAGGTCAACCTCTACACCATCTCCTACG
TGGTATACGTTCCCATTACATAAAACTGGTGATAATTCTGTTGGGTCAAGCCGATCAGCC
TACGAATTTTTGATCACAAATTCAGATGAGAAAGAAAATGGTATTCCTGTCAAATTAACA
ATAGAAAATCACGAAGTAAATTCTCTGCATGGAGACGGGGGCGAGCAAATTGCCAAGAAA
ATTGTCATACCAGATAAATTTATCCAGCATATCTAG
[0470] Further information on DER3 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000005373.
[0471] It will be appreciated that, by "DER3", we include fragments
or variants thereof having equivalent DER3-like activity.
[0472] HRD3 is another S. cerevisiae helper protein of interest for
the present invention. Hrd3p is a resident protein of the ER
membrane that plays a central role in ER-associated protein
degradation (ERAD). It forms an HRD complex with Hrd1p and ERAD
determinants that engage in lumen to cytosol communication and
coordination of ERAD events. A published protein sequence for the
protein Hrd3p is as follows:
TABLE-US-00054
MITLLLYLCVICNAIVLIRADSIADPWPEARHLLNTIAKSRDPMKEAAMEPNADEFVGFY
VPMDYSPRNEEKNYQSIWQNEITDSQRHIYELLVQSSEQFNNSEATYTLSQIHLWSQYNF
PHNMTLAHKYLEKFNDLTHFTNHSAIFDLAVMYATGGCASGNDQTVIPQDSAKALLYYQR
AAQLGNLKAKQVLAYKYYSGFNVPRNFHKSLVLYRDIAEQLRKSYSRDEWDIVFPYWESY
NVRISDFESGLLGKGLNSVPSSTVRKRTTRPDIGSPFIAQVNGVQMTLQIEPMGRFAFNG
NDGNINGDEDDEDASERRIIRIYYAALNDYKGTYSQSRNCERAKNLLELTYKEFQPHVDN
LDPLQVFYYVRCLQLLGHMYFTGEGSSKPNIHMAEEILTTSLEISRRAQGPIGRACIDLG
LINQYITNNISQAISYYMKAMKTQANNGIVEFQLSKLATSFPEEKIGDPFNLMETAYLNG
FIPAIYEFAVMIESGMNSKSSVENTAYLFKTFVDKNEAIMAPKLRTAFAALINDRSEVAL
WAYSQLAEQGYETAQVSAAYLMYQLPYEFEDPPRTTDQRKTLAISYYTRAFKQGNIDAGV
VAGDIYFQMQNYSKAMALYQGAALKYSIQAIWNLGYMHEHGLGVNRDFHLAKRYYDQVSE
HDHRFYLASKLSVLKLHLKSWLTWITREKVNYWKPSSPLNPNEDTQHSKTSWYKQLTKIL
QRMRHKEDSDKAAEDSHKHRTVVQNGANHRGDDQEEASEILGFQMEDLVTMGCILGIFLL
SILMSTLAARRGWNVRFNGAQLNANGNRQQEQQQQQQAQGPPGWDFNVQIFAI*
[0473] HRD3 is encoded by a non-essential gene comprising an ORF
that is 2.502 kbp in size and is located on chromosome XII. A
published nucleotide coding sequence of HRD3 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00055
ATGATAACACTCTTATTATACCTGTGCGTAATATGTAACGCAATAGTGTTAATAAGGGCT
GATTCGATAGCGGACCCTTGGCCTGAAGCGCGACATCTACTAAATACCATAGCTAAGTCC
AGAGACCCAATGAAAGAAGCTGCTATGGAACCCAATGCAGATGAATTTGTTGGATTCTAT
GTACCGATGGATTATTCCCCACGTAATGAGGAAAAAAACTACCAGAGCATTTGGCAAAAC
GAAATCACAGATTCTCAACGTCATATTTATGAATTACTTGTACAATCAAGTGAACAATTC
AACAACTCAGAAGCAACATATACACTTAGCCAGATTCACCTTTGGAGTCAATATAATTTC
CCGCATAATATGACTTTGGCACACAAATACTTAGAAAAATTCAATGATCTAACCCACTTC
ACCAATCATTCGGCCATCTTCGACTTAGCTGTGATGTATGCCACTGGGGGATGTGCTTCT
GGTAATGATCAAACCGTGATCCCTCAGGATTCTGCTAAAGCACTGCTATATTACCAAAGG
GCTGCCCAACTAGGGAATTTAAAGGCTAAGCAAGTGCTAGCTTATAAATACTATTCTGGC
TTCAATGTCCCACGAAATTTTCATAAATCTTTAGTATTGTACAGGGACATTGCTGAACAG
CTGAGAAAGTCGTACTCCAGGGACGAATGGGATATTGTCTTCCCCTATTGGGAAAGTTAC
AACGTGAGAATATCGGATTTTGAGAGTGGCCTATTAGGTAAAGGTTTGAATTCCGTTCCA
TCTTCTACAGTAAGGAAAAGAACTACGAGACCAGATATTGGTTCACCCTTTATTGCGCAA
GTTAACGGTGTACAGATGACCTTGCAAATCGAACCGATGGGTAGGTTCGCTTTCAACGGT
AACGATGGCAACATAAATGGCGACGAAGATGACGAGGATGCCAGTGAAAGACGAATCATT
CGGATATATTATGCAGCTTTGAATGATTATAAAGGAACATATTCACAAAGCAGAAATTGT
GAGCGCGCCAAAAACTTGTTGGAATTAACGTACAAGGAATTTCAGCCTCATGTCGACAAT
TTGGATCCTTTGCAAGTATTTTACTACGTCCGTTGCTTACAATTATTGGGGCACATGTAT
TTCACCGGCGAAGGCTCCTCGAAGCCTAATATTCATATGGCCGAAGAGATCCTGACCACG
TCGCTAGAAATAAGCAGAAGGGCACAGGGACCTATAGGTAGAGCGTGCATAGATCTGGGC
TTAATAAATCAATACATCACAAACAATATTTCTCAAGCAATTTCGTATTATATGAAAGCT
ATGAAAACACAAGCTAACAATGGAATCGTAGAATTCCAATTATCCAAATTGGCCACTTCA
TTCCCTGAAGAAAAAATCGGCGACCCATTTAACTTAATGGAAACTGCCTACTTGAATGGA
TTCATTCCAGCCATATATGAGTTTGCAGTAATGATCGAATCTGGAATGAACAGTAAGAGT
AGTGTGGAAAACACTGCTTACCTGTTCAAAACATTCGTTGACAAAAACGAAGCTATTATG
GCACCTAAACTGAGGACAGCATTTGCCGCATTAATCAACGATCGTTCAGAAGTGGCTTTA
TGGGCTTATTCCCAACTAGCCGAGCAAGGCTACGAGACTGCTCAAGTCTCTGCCGCCTAC
TTAATGTACCAGTTGCCATATGAGTTTGAGGATCCTCCAAGAACCACAGATCAGAGAAAA
ACTTTGGCAATTTCCTACTATACAAGAGCGTTTAAACAGGGAAATATAGATGCTGGTGTT
GTCGCGGGAGATATCTATTTTCAGATGCAGAATTACAGTAAAGCTATGGCTCTTTATCAG
GGTGCAGCTTTGAAGTACTCTATACAGGCTATCTGGAACTTAGGGTACATGCATGAGCAT
GGGCTAGGTGTAAACAGAGATTTCCATCTTGCTAAACGTTACTACGACCAAGTTTCAGAA
CACGATCATAGATTTTACTTGGCTTCCAAATTGAGTGTTTTAAAATTACACCTAAAGTCA
TGGTTGACTTGGATCACCAGAGAAAAAGTAAACTACTGGAAACCTTCCTCGCCACTTAAC
CCTAACGAAGATACTCAGCACTCGAAGACTTCATGGTACAAGCAATTGACGAAGATTCTA
CAAAGAATGAGACATAAGGAGGATAGTGACAAAGCTGCGGAAGATTCTCACAAACACAGA
ACTGTAGTGCAGAATGGAGCTAACCATAGGGGTGACGACCAAGAGGAGGCTTCCGAGATT
TTGGGCTTCCAAATGGAGGATCTTGTTACGATGGGATGTATCTTGGGGATATTCCTATTA
AGTATATTAATGAGTACACTGGCGGCCCGTAGAGGCTGGAATGTCCGTTTCAATGGAGCA
CAATTAAATGCAAATGGTAACCGGCAGCAAGAGCAACAACAACAACAACAAGCACAAGGT
CCCCCGGGCTGGGACTTCAATGTTCAGATATTCGCCATATGA
[0474] Further information on HRD3 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000004197.
[0475] It will be appreciated that, by "HRD3", we include fragments
or variants thereof having equivalent HRD3-like activity.
[0476] UBC7 is another S. cerevisiae helper protein of interest for
the present invention and is also known as QRI8. Ubc7p is a
ubiquitin conjugating enzyme, involved in the ER-associated protein
degradation pathway. It requires Cue1p for recruitment to the ER
membrane and is proposed to be involved in chromatin assembly. A
published protein sequence for the protein Ubc7p is as follows:
TABLE-US-00056 MSKTAQKRLLKELQQLIKDSPPGIVAGPKSENNIFIWDCLIQGPPDTP
YADGVFNAKLEFPKDYPLSPPKLTFTPSILHPNIYPNGEVCISILHSPG
DDPNMYELAEERWSPVQSVEKILLSVMSMLSEPNIESGANIDACILW
RDNRPEFERQVKLSILKSLGF*
[0477] UBC7 is encoded by a non-essential gene comprising an ORF
that is 0.498 kbp in size and is located on chromosome XIII. A
published nucleotide coding sequence of UBC7 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00057 ATGTCGAAAACCGCTCAGAAACGTCTCCTCAAGGAGCTTCAACAGTTA
ATTAAAGATTCTCCACCTGGTATAGTGGCTGGTCCCAAATCGGAGAATA
ACATATTCATTTGGGACTGCCTAATTCAAGGGCCTCCAGATACGCCATA
CGCTGATGGTGTTTTTAATGCTAAGCTAGAGTTTCCTAAAGACTATCCGT
TATCTCCACCTAAACTTACTTTCACACCCAGCATACTACATCCAAATATT
TATCCAAATGGGGAAGTGTGCATATCCATTCTACACTCCCCTGGTGATG
ATCCTAACATGTACGAATTAGCGGAAGAAAGATGGTCGCCAGTGCAAA
GTGTAGAAAAAATTCTATTAAGTGTTATGAGCATGTTGAGTGAGCCCAAT
ATCGAAAGTGGTGCCAACATTGATGCTTGCATCTTGTGGAGAGATAATA
GACCTGAATTTGAGAGACAGGTAAAGTTATCCATTTTGAAATCATTAGGA TTCTGA
[0478] Further information on UBC7 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000004624.
[0479] It will be appreciated that, by "UBC7", we include fragments
or variants thereof having equivalent UBC7-like activity.
[0480] DOA4 is another S. cerevisiae helper protein of interest for
the present invention and is also known as DOS1, MUT4, NPI2, SSV7,
and UBP4. Doa4p is a ubiquitin hydrolase, required for recycling
ubiquitin from proteasome-bound ubiquitinated intermediates, which
acts at the late endosome/prevacuolar compartment to recover
ubiquitin from ubiquitinated membrane proteins en route to the
vacuole. A published protein sequence for the protein Doa4p is as
follows:
TABLE-US-00058
MEQNIISTIRDECIRHRSKYLTIAQLTAIAEAKINEFIITGKAKDQDLSSLLDKCIDILS
IYKKNSKDIKNIISCKNKGAMISSNSVMIIQLNYVYYKVIHIIVTTNIPHLSEFAKIKLH
KSTSDEGNGNNNNNEFQLMNIYNTLLETLLKDENIAKIKSFIKSSIKQTKLNHEQEECNL
MRTGSYITSNQLNSLISSSANSASSQMEILLIDIRSRLEFNKSHIDTKNIICLEPISFKM
SYSDHDLEKKSLITSPNSEIKMFQSRNLFKFIILYTDANEYNVKQQSVLLDILVNHSFEK
PISDDFTKIFILESGFPGWLKSNYGRQVSSSFPSNNNIKDDSVYINGNTSGLSLQHLPKM
SPSIRHSMDDSMKEMLVAPTPLNHLQQQQQQQSDNDHVLKRSSSFKKLFSNYTSPNPKNS
NSNLYSISSLSISSSPSPLPLHSPDPVKGNSLPINYPETPHLWKNSETDFMTNQREQLNH
NSFAHIAPINTKAITSPSRTATPKLQRFPQTISMNLNMNSNGHSSATSTIQPSCLSLSNN
DSLDHTDVTPTSSHNYDLDFAVGLENLGNSCYMNCIIQCILGTHELTQIFLDDSYAKHIN
INSKLGSKGILAKYFARLVHMMYKEQVDGSKKISISPIKFKLACGSVNSLFKTASQQDCQ
EFCQFLLDGLHEDLNQCGSNPPLKELSQEAEARREKLSLRIASSIEWERFLTTDFSVIVD
LFQGQYASRLKCKVCSHTSTTYQPFTVLSIPIPKKNSRNNITIEDCFREFTKCENLEVDE
QWLCPHCEKRQPSTKQLTITRLPRNLIVHLKRFDNLLNKNNDFVIYPFLLDLTPFWANDF
DGVFPPGVNDDELPIRGQIPPFKYELYGVACHFGTLYGGHYTAYVKKGLKKGWLYFDDTK
YKPVKNKADAINSNAYVLFYHRVYGV*
[0481] DOA4 is encoded by a non-essential gene comprising an ORF
that is 2.781 kbp in size and is located on chromosome IV. A
published nucleotide coding sequence of DOA4 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00059
ATGGAGCAGAATATTATTAGTACCATAAGGGATGAGTGTATTCGTCACCGGTCGAAGTAC
CTTACGATAGCACAACTAACCGCTATTGCAGAGGCTAAAATTAACGAATTCATCATAACT
GGTAAGGCAAAAGATCAAGATTTGAGCAGTCTTCTAGATAAATGCATCGATATTTTATCT
ATTTACAAGAAGAACTCGAAAGATATCAAAAATATTATATCGTGCAAAAATAAGGGTGCA
ATGATTAGTTCAAATTCCGTAATGATTATTCAATTAAATTATGTTTACTACAAGGTAATT
CACATTATTGTAACAACCAATATTCCTCATTTAAGTGAATTCGCCAAGATTAAATTACAT
AAGAGCACGAGTGATGAGGGCAACGGTAATAACAACAATAATGAATTTCAACTCATGAAC
ATTTACAACACTTTGCTGGAAACCTTATTAAAAGATGAAAACATTGCAAAAATAAAAAGT
TTCATTAAGTCTTCCATAAAACAAACAAAATTGAACCATGAGCAAGAAGAATGTAACCTG
ATGAGAACGGGTTCCTATATCACTTCCAATCAATTAAACTCCCTAATAAGTTCATCAGCA
AATTCTGCTTCCTCCCAAATGGAGATACTACTGATAGATATACGATCAAGGTTGGAATTC
AACAAGTCACATATTGATACAAAAAATATTATATGCCTGGAGCCTATTTCTTTTAAAATG
TCATATTCAGATCATGATTTGGAGAAAAAATCATTAATTACTTCTCCTAATAGTGAGATT
AAAATGTTTCAAAGTAGAAATCTTTTCAAGTTTATCATTCTCTATACAGACGCAAACGAA
TACAATGTTAAACAGCAGTCTGTCCTGTTGGACATTCTGGTGAATCATTCCTTTGAAAAA
CCAATATCCGATGACTTTACCAAAATTTTCATTCTGGAATCTGGTTTTCCAGGTTGGCTT
AAGTCAAATTATGGGAGGCAAGTATCATCATCTTTTCCATCAAATAACAATATTAAAGAT
GATAGTGTTTATATTAATGGTAACACTTCTGGCCTAAGTTTACAACATTTACCTAAGATG
TCTCCCAGTATAAGACATTCAATGGACGACTCTATGAAAGAAATGCTAGTTGCGCCTACT
CCATTAAATCATCTTCAACAACAGCAACAACAGCAATCAGACAATGATCATGTGCTAAAA
AGATCTTCAAGTTTCAAAAAATTATTCTCAAATTATACGTCTCCTAATCCGAAGAATTCA
AATTCAAACTTATATTCTATATCTTCGTTGTCCATATCTAGTTCACCATCGCCTTTACCT
CTACATTCGCCTGACCCAGTTAAGGGCAATTCATTGCCAATCAATTATCCGGAAACGCCA
CATCTTTGGAAAAACAGTGAGACAGATTTTATGACAAATCAAAGAGAACAGTTGAATCAC
AACTCTTTTGCTCACATAGCTCCTATCAACACGAAGGCCATCACTTCTCCATCAAGAACT
GCCACACCGAAGTTACAACGCTTCCCGCAAACAATTAGTATGAACCTTAATATGAACTCC
AATGGACACAGTTCTGCCACCTCTACCATTCAACCTTCGTGTCTATCCTTGTCTAATAAT
GACTCTTTAGATCATACAGATGTTACACCAACTTCTTCTCATAATTATGACCTTGATTTC
GCGGTTGGTTTGGAAAATCTAGGAAATTCGTGTTACATGAACTGTATCATTCAGTGTATC
TTAGGTACACACGAATTAACCCAAATCTTTTTGGACGATTCATATGCTAAACACATCAAT
ATTAATAGTAAGTTGGGATCGAAAGGTATTCTGGCAAAATATTTTGCAAGGTTGGTTCAT
ATGATGTATAAGGAACAGGTTGATGGTTCAAAGAAAATTTCCATATCACCGATAAAATTT
AAATTGGCATGTGGATCTGTTAACTCATTATTTAAGACTGCATCCCAACAGGACTGCCAA
GAGTTTTGCCAATTCCTTCTAGATGGTCTTCATGAAGACTTGAACCAATGCGGTTCAAAC
CCACCTTTGAAGGAGCTTTCTCAAGAAGCTGAGGCGAGAAGAGAAAAACTGTCTTTGCGA
ATTGCCTCGTCAATTGAGTGGGAACGATTCTTGACTACTGATTTCAGTGTTATTGTCGAC
TTATTTCAGGGACAATACGCCTCACGACTAAAATGTAAAGTCTGTAGTCATACCTCGACA
ACATACCAACCTTTTACGGTGCTGTCAATCCCTATTCCTAAAAAAAATTCCCGAAATAAT
ATTACCATTGAAGATTGTTTCAGAGAGTTCACCAAATGTGAGAACTTGGAAGTGGATGAG
CAATGGTTGTGCCCACATTGTGAAAAAAGGCAGCCCTCCACGAAACAATTGACAATAACG
AGATTACCGAGGAATCTGATAGTCCATTTAAAGAGATTTGATAATTTATTAAACAAAAAT
AATGACTTCGTCATATACCCTTTTTTGTTGGACTTGACTCCATTTTGGGCCAATGATTTT
GACGGGGTTTTTCCTCCAGGTGTTAATGACGATGAACTACCAATAAGGGGACAAATACCA
CCTTTTAAGTATGAATTATATGGTGTAGCATGCCACTTTGGTACTTTGTATGGTGGTCAT
TATACAGCCTATGTGAAAAAGGGATTAAAGAAGGGATGGCTATATTTTGATGATACCAAA
TATAAACCTGTCAAAAACAAAGCCGATGCAATTAACTCTAATGCATACGTTTTGTTTTAT
CACCGCGTCTACGGTGTTTGA
[0482] Further information on DOA4 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000002476.
[0483] It will be appreciated that, by "DOA4", we include fragments
or variants thereof having equivalent DOA4-like activity.
[0484] HAC1 is another S. cerevisiae helper protein of interest for
the present invention, and is also known as ERN4 and IRE15. Hac1p,
is a bZIP transcription factor (ATF/CREB1 homolog) that regulates
the unfolded protein response, via UPRE binding, and membrane
biogenesis. ER stress-induced splicing pathway utilising Ire1p,
Trl1p and Ada5p facilitates efficient Hac1p synthesis. A published
protein sequence for the protein Hac1p is as follows:
TABLE-US-00060 MEMTDFELTSNSQSNLAIPTNFKSTLPPRKRAKTKEEKEQRRIERILRNR
RAAHQSREKKRLHLQYLERKCSLLENLLNSVNLEKLADHEDALTCSHD
AFVASLDEYRDFQSTRGASLDTRASSHSSSDTFTPSPLNCTMEPATLS
PKSMRDSASDQETSWELQMFKTENVPESTTLPAVDNNNLFDAVASPL
ADPLCDDIAGNSLPFDNSIDLDNWRNPEAQSGLNSFELNDFFITS*
[0485] HAC1 is encoded by a non-essential gene that is located on
chromosome VI. A published nucleotide coding sequence of HAC1, that
has been processed to remove introns, is 0.717 kbp in size and is
as follows (although it will be appreciated that the sequence can
be modified by degenerate substitutions to obtain alternative
nucleotide sequences which encode an identical protein
product):
TABLE-US-00061
ATGGAAATGACTGATTTTGAACTAACTAGTAATTCGCAATCGAACTTGGCTATCCCTACC
AACTTCAAGTCGACTCTGCCTCCAAGGAAAAGAGCCAAGACAAAAGAGGAAAAGGAACAG
CGAAGGATCGAGCGTATTTTGAGAAACAGAAGAGCTGCTCACCAGAGCAGAGAGAAAAAA
AGACTACATCTGCAGTATCTCGAGAGAAAATGTTCTCTTTTGGAAAATTTACTGAACAGC
GTCAACCTTGAAAAACTGGCTGACCACGAAGACGCGTTGACTTGCAGCCACGACGCTTTT
GTTGCTTCTCTTGACGAGTACAGGGATTTCCAGAGCACGAGGGGCGCTTCACTGGACACC
AGGGCCAGTTCGCACTCGTCGTCTGATACGTTCACACCTTCACCTCTGAACTGTACAATG
GAGCCTGCGACTTTGTCGCCCAAGAGTATGCGCGATTCCGCGTCGGACCAAGAGACTTCA
TGGGAGCTGCAGATGTTTAAGACGGAAAATGTACCAGAGTCGACGACGCTACCTGCCGTA
GACAACAACAATTTGTTTGATGCGGTGGCCTCGCCGTTGGCAGACCCACTCTGCGACGAT
ATAGCGGGAAACAGTCTACCCTTTGACAATTCAATTGATCTTGACAATTGGCGTAATCCA
GAAGCGCAGTCAGGTTTGAATTCATTTGAATTGAATGATTTCTTCATCACTTCATGA
[0486] Further information on HAC1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000001863.
[0487] It will be appreciated that, by "HAC1", we include fragments
or variants thereof having equivalent HAC1-like activity.
[0488] SEC63 is another S. cerevisiae helper protein of interest
for the present invention. It is also known as PTL1. It is an
essential subunit of the Sec63 complex (Sec63p, Sec62p, Sec66p and
Sec72p); with Sec61 complex, Kar2p/BiP and Lhs1p it forms a channel
competent for SRP-dependent and post-translational SRP-independent
protein targeting and import into the ER. A published protein
sequence for the protein Sec63p is as follows:
TABLE-US-00062
MPTNYEYDEASETWPSFILTGLLMVVGPMTLLQIYQIFFGANAEDGNSGKSKEFNEEVFK
NLNEEYTSDEIKQFRRKFDKNSNKKSKIWSRRNIIIIVGWILVAILLQRINSNDAIKDAA
TKLFDPYEILGISTSASDRDIKSAYRKLSVKFHPDKLAKGLTPDEKSVMEETYVQITKAY
ESLTDELVRQNYLKYGHPDGPQSTSHGIALPRFLVDGSASPLLVVCYVALLGLILPYFVS
RWWARTQSYTKKGIHNVTASNFVSNLVNYKPSEIVTTDLILHWLSFAHEFKQFFPDLQPT
DFEKLLQDHINRRDSGKLNNAKFRIVAKCHSLLHGLLDIACGFRNLDIALGAINTFKCIV
QAVPLTPNCQILQLPNVDKEHFITKTGDIHTLGKLFTLEDAKIGEVLGIKDQAKLNETLR
VASHIPNLKIIKADFLVPGENQVTPSSTPYISLKVLVRSAKQPLIPTSLIPEENLTEPQD
FESQRDPFAMMSKQPLVPYSFAPFFPTKRRGSWCCLVSSQKDGKILQTPIIIEKLSYKNL
NDDKDFFDKRIKMDLTKHEKFDINDWEIGTIKIPLGQPAPETVGDFFFRVIVKSTDYFTT
DLDITMNMKVRDSPAVEQVEVYSEEDDEYSTDDDETESDDESDASDYTDIDTDTEAEDDE
SPE*
[0489] SEC63 is encoded by an essential gene comprising an ORF that
is 1.192 kbp in size and is located on chromosome XV. A published
nucleotide coding sequence of SEC63 is as follows, although it will
be appreciated that the sequence can be modified by degenerate
substitutions to obtain alternative nucleotide sequences which
encode an identical protein product:
TABLE-US-00063
ATGCCTACAAATTACGAGTATGATGAGGCTAGTGAGACGTGGCCGTCCTTCATTTTAACG
GGGCTCTTGATGGTCGTCGGGCCTATGACACTGCTTCAAATATACCAAATTTTTTTTGGG
GCCAATGCTGAAGATGGGAATTCAGGGAAGAGTAAGGAGTTTAATGAGGAAGTTTTCAAG
AACTTGAATGAAGAATACACCAGTGATGAAATCAAACAATTTAGAAGGAAGTTTGATAAA
AATAGTAATAAGAAGTCCAAAATATGGAGCAGGAGAAATATTATAATTATTGTGGGTTGG
ATCTTAGTTGCAATTCTTCTGCAAAGGATTAATAGTAATGACGCGATTAAAGACGCTGCT
ACAAAATTATTTGATCCTTATGAAATCCTTGGTATCTCTACTAGTGCTTCCGATAGAGAC
ATCAAATCTGCTTATAGAAAATTATCTGTTAAATTTCATCCAGATAAATTAGCAAAGGGC
CTAACACCTGATGAGAAAAGTGTGATGGAAGAAACTTATGTTCAGATTACGAAGGCTTAC
GAATCCCTTACTGACGAATTGGTTAGGCAAAACTATTTGAAATACGGTCATCCAGATGGC
CCACAATCTACTTCACATGGTATCGCTCTACCAAGATTTTTGGTAGATGGAAGTGCATCT
CCATTATTAGTGGTTTGTTATGTTGCGCTACTAGGTTTAATCTTGCCATATTTTGTTAGT
AGATGGTGGGCAAGAACACAATCGTATACTAAGAAGGGAATACATAATGTGACGGCTTCT
AATTTTGTTAGTAACTTAGTCAATTACAAGCCATCTGAGATTGTCACCACAGATTTGATC
TTACACTGGTTATCATTTGCTCATGAATTTAAACAATTCTTCCCGGATTTGCAACCAACG
GATTTTGAAAAACTTTTGCAAGATCATATTAACCGCAGAGATAGTGGTAAACTTAACAAT
GCGAAATTTAGAATAGTGGCCAAATGTCACTCTTTGTTACACGGTTTATTGGATATTGCT
TGTGGATTCAGAAATTTAGATATTGCATTGGGTGCAATCAATACTTTCAAGTGTATTGTT
CAGGCTGTACCATTAACACCAAACTGTCAAATCCTTCAATTGCCGAACGTAGATAAAGAG
CACTTTATTACCAAAACCGGAGATATTCATACATTAGGTAAATTGTTTACTTTAGAAGAT
GCCAAGATTGGTGAGGTTCTTGGAATAAAGGATCAAGCAAAGTTAAACGAAACTTTGAGA
GTTGCATCGCATATTCCAAATCTAAAGATCATCAAGGCAGACTTCCTTGTCCCAGGTGAG
AACCAAGTAACACCATCATCTACCCCATACATTTCTTTGAAAGTACTGGTTCGTTCTGCT
AAACAGCCATTGATACCAACTAGCTTAATTCCTGAAGAAAATTTAACAGAACCTCAAGAT
TTTGAATCTCAAAGAGATCCATTTGCTATGATGAGTAAACAGCCACTCGTCCCATATTCC
TTTGCACCATTTTTCCCTACAAAGAGACGTGGGAGTTGGTGCTGTCTGGTAAGTTCTCAA
AAAGATGGTAAAATACTTCAAACGCCAATTATCATTGAAAAGCTATCTTACAAGAACTTG
AACGATGACAAAGATTTCTTTGATAAGAGGATAAAAATGGATTTAACCAAACACGAAAAA
TTCGATATAAATGATTGGGAAATCGGGACCATAAAAATTCCATTAGGTCAGCCTGCACCT
GAAACTGTTGGTGATTTCTTTTTTAGAGTAATCGTTAAATCCACAGATTATTTCACTACA
GATTTGGATATTACCATGAATATGAAAGTTCGTGATTCTCCTGCAGTGGAACAAGTAGAG
GTGTATTCTGAGGAGGATGATGAGTACTCTACTGATGACGACGAAACCGAAAGTGATGAT
GAAAGTGATGCTAGCGATTATACTGATATCGATACGGATACAGAAGCTGAAGATGATGAA
TCACCAGAATAG
[0490] Further information on SEC63 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000005780
[0491] It will be appreciated that, by "SEC63", we include
fragments or variants thereof having equivalent SEC63-like
activity.
[0492] YDJ1 is another S. cerevisiae helper protein of interest for
the present invention. It is also known as MASS and HSP40. It is a
protein chaperone involved in regulation of the HSP90 and HSP70
functions; involved in protein translocation across membranes;
member of the DnaJ family, and is located in the cytoplasm. A
published protein sequence for the protein Ydj1p is as follows:
TABLE-US-00064 MVKETKFYDILGVPVTATDVEIKKAYRKCALKYHPDKNPSEEAAEKFK
EASAAYEILSDPEKRDIYDQFGEDGLSGAGGAGGFPGGGFGFGDDIF
SQFFGAGGAQRPRGPQRGKDIKHEISASLEELYKGRTAKLALNKQILC
KECEGRGGKKGAVKKCTSCNGQGIKFVTRQMGPMIQRFQTECDVCH
GTGDIIDPKDRCKSCNGKKVENERKILEVHVEPGMKDGQRIVFKGEAD
QAPDVIPGDVVFIVSERPHKSFKRDGDDLVYEAEIDLLTAIAGGEFALE
HVSGDWLKVGIVPGEVIAPGMRKVIEGKGMPIPKYGGYGNLIIKFTIKFP
ENHFTSEENLKKLEEILPPRIVPAIPKKATVDECVLADFDPAKYNRTRA
SRGGANYDSDEEEQGGEGVQCASQ*
[0493] YDJ1 is encoded by a non-essential gene comprising an ORF
that is 1.230 kbp in size and is located on chromosome XIV. A
published nucleotide coding sequence of YDJ1 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00065
ATGGTTAAAGAAACTAAGTTTTACGATATTCTAGGTGTTCCAGTAACTGCCACTGATGTC
GAAATTAAGAAAGCTTATAGAAAATGCGCCTTAAAATACCATCCAGATAAGAATCCAAGT
GAGGAAGCTGCAGAAAAGTTCAAAGAAGCTTCAGCAGCCTATGAAATTTTATCAGATCCT
GAAAAGAGAGATATATATGACCAATTTGGTGAAGATGGTCTAAGTGGTGCTGGTGGCGCT
GGCGGATTCCCAGGTGGTGGATTCGGTTTTGGTGACGATATCTTTTCCCAATTCTTTGGT
GCTGGTGGCGCACAAAGACCAAGAGGTCCCCAAAGAGGTAAAGATATCAAGCATGAAATT
TCTGCCTCACTTGAAGAATTATATAAGGGTAGGACAGCTAAGTTAGCCCTTAACAAACAG
ATCCTATGTAAAGAATGTGAAGGTCGTGGTGGTAAGAAAGGCGCCGTCAAGAAGTGTACC
AGCTGTAATGGTCAAGGTATTAAATTTGTAACAAGACAAATGGGTCCAATGATCCAAAGA
TTCCAAACAGAGTGTGATGTCTGTCACGGTACTGGTGATATCATTGATCCTAAGGATCGT
TGTAAATCTTGTAACGGTAAGAAAGTTGAAAACGAAAGGAAGATCCTAGAAGTCCATGTC
GAACCAGGTATGAAAGATGGTCAAAGAATCGTTTTCAAAGGTGAAGCTGACCAAGCCCCA
GATGTCATTCCAGGTGATGTTGTCTTCATAGTTTCTGAGAGACCACACAAGAGCTTCAAG
AGAGATGGTGATGATTTAGTATATGAGGCTGAAATTGATCTATTGACTGCTATCGCTGGT
GGTGAATTTGCATTGGAACATGTTTCTGGTGATTGGTTAAAGGTCGGTATTGTTCCAGGT
GAAGTTATTGCCCCAGGTATGCGTAAGGTCATCGAAGGTAAAGGTATGCCAATTCCAAAA
TACGGTGGCTATGGTAATTTAATCATCAAATTTACTATCAAGTTCCCAGAAAACCATTTC
ACATCAGAAGAAAACTTGAAGAAGTTAGAAGAAATTTTGCCTCCAAGAATTGTCCCAGCC
ATTCCAAAGAAAGCTACTGTGGACGAATGTGTACTCGCAGACTTTGACCCAGCCAAATAC
AACAGAACACGGGCCTCCAGGGGTGGTGCAAACTATGATTCCGATGAAGAAGAACAAGGT
GGCGAAGGTGTTCAATGTGCATCTCAATGA
[0494] Further information on YDJ1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000005008
[0495] It will be appreciated that, by "YDJ1", we include fragments
or variants thereof having equivalent YDJ1-like activity.
[0496] XDJ1 is another S. cerevisiae helper protein of interest for
the present invention. It is a putative chaperone, a homolog of E.
coli DnaJ, and is closely related to Ydj1p. A published protein
sequence for the protein Xdj1p is as follows:
TABLE-US-00066 MSGSDRGDRLYDVLGVTRDATVQEIKTAYRKLALKHHPDKYVDQDSKEVN
EIKFKEITAAYEILSDPEKKSHYDLYGDDNGAASSGGANGFGDEDFMNFF
NNFFNNGSHDGNNFPGEYDAYEEGNSTSSKDIDIDISLTLKDLYMGKKLK
FDLKRQVICIKCHGSGWKPKRKIHVTHDVECESCAGKGSKERLKRFGPGL
VASQWVVCEKCNGKGKYTKRPKNPKNFCPDCAGLGLLSKKEIITVNVAPG
HHFNDVITVKGMADEEIDKTTCGDLKFHLTEKQENLEQKQIFLKNFDDGA
GEDLYTSITISLSEALTGFEKFLTKTFDDRLLTLSVKPGRVVRPGDTIKI
ANEGWPILDNPHGRCGDLYVFVHIEFPPDNWFNEKSELLAIKTNLPSSSS
CASHATVNTEDDSNLTNNETISNFRIIHTDDLPEGIRPFKPEAQDSAHQK ARSSYCCIQ*
[0497] XDJ1 is encoded by a non-essential gene comprising an ORF
that is 1.380 kbp in size and is located on chromosome XII. A
published nucleotide coding sequence of XDJ1 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00067
ATGAGTGGCAGTGATAGAGGAGACCGGTTATACGATGTGTTGGGGGTGACGAGAGATGCG
ACCGTGCAAGAGATTAAAACTGCTTACAGAAAGCTTGCCCTGAAACATCATCCGGACAAG
TATGTGGATCAAGACTCAAAGGAGGTAAATGAAATCAAATTCAAAGAGATCACTGCCGCT
TACGAGATCTTGAGCGATCCGGAGAAGAAATCACATTACGACTTGTATGGTGATGATAAT
GGTGCCGCTAGCAGCGGTGGCGCTAATGGCTTTGGAGATGAAGATTTTATGAACTTCTTT
AACAATTTCTTCAATAATGGAAGTCACGATGGAAATAATTTCCCTGGCGAGTATGATGCG
TACGAAGAGGGCAACTCTACAAGCTCTAAGGATATCGATATCGATATATCTCTTACTTTG
AAGGATTTGTACATGGGCAAGAAGCTGAAGTTTGATTTAAAGAGACAGGTCATCTGTATA
AAGTGCCACGGTTCTGGCTGGAAACCAAAGAGGAAAATTCACGTTACACACGATGTGGAA
TGTGAATCATGCGCTGGAAAGGGTTCAAAGGAACGTCTGAAGAGGTTTGGTCCCGGTTTG
GTAGCTTCGCAATGGGTGGTCTGTGAGAAATGTAATGGTAAGGGGAAGTACACTAAAAGA
CCCAAGAATCCAAAGAACTTTTGCCCCGATTGCGCAGGCTTGGGGCTCCTGTCAAAGAAG
GAAATCATCACAGTGAACGTGGCTCCGGGACACCACTTTAACGACGTAATTACAGTCAAG
GGGATGGCGGACGAGGAAATCGATAAGACCACATGTGGTGATTTAAAGTTCCATCTCACT
GAAAAACAAGAAAACTTGGAGCAGAAGCAAATCTTTTTGAAGAACTTTGACGACGGCGCC
GGGGAAGATTTGTATACAAGCATTACCATATCGTTAAGCGAGGCCTTGACGGGATTTGAG
AAATTTTTGACAAAAACCTTCGACGACAGGTTACTAACATTGAGCGTTAAACCTGGCAGA
GTAGTAAGACCTGGTGACACCATCAAAATCGCCAATGAAGGTTGGCCCATTTTAGATAAC
CCTCATGGCCGGTGCGGCGATCTGTATGTTTTCGTTCATATTGAATTTCCACCAGATAAC
TGGTTCAATGAAAAATCAGAACTACTAGCAATAAAAACGAATCTGCCGTCATCTTCATCT
TGTGCCTCACATGCGACTGTAAATACTGAAGATGACAGCAACCTGACTAACAACGAAACT
ATATCAAATTTCCGGATCATTCACACGGACGATCTTCCAGAAGGGATAAGGCCGTTCAAG
CCAGAAGCACAGGATTCAGCGCATCAGAAAGCAAGAAGTTCGTACTGCTGTATCCAATGA
[0498] Further information on XDJ1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000004080
[0499] It will be appreciated that, by "XDJ1", we include fragments
or variants thereof having equivalent XDJ1-like activity.
[0500] APJ1 is another S. cerevisiae helper protein of interest for
the present invention. It is a putative chaperone of the HSP40
(DnaJ) family; over expression of which interferes with propagation
of the [Psi+] prion. A published protein sequence for the protein
Apj1p is as follows:
TABLE-US-00068
MQQNTSLYDSLNVTAAASTSEIKKAYRNAALKYHPDKNNHTEESKRKFQEICQAYEILKD
NRLRALYDQYGTTDEVLIQEQQAQAQRQQAGPFSSSSNFDTEAMSFPDLSPGDLFAQFFN
SSATPSSNGSKSSFNFSFNNSSTPSFSFVNGSGVNNLYSSSAKYNSNDEDHHLDRGPDIK
HNLKCTLKELYMGKTAKLGLNRTRICSVCDGHGGLKKCTCKTCKGQGIQTQTRRMGPLVQ
SWSQTCADCGGAGVFVKNKDICQQCQGLGFIKERKILQVTVQPGSCHNQLIVLTGEGDEV
ISTKGGGHEKVIPGDVVITILRLKDPNFQVINYSNLICKKCKIDFMTSLCGGVVYIEGHP
SGKLIKLDIIPGEILKPGCFKTVEDMGMPKFINGVRSGFGHLYVKFDVTYPERLEPENAK
KIQNILANDKYIKAERSTMETADSDCYCDLEKSYDSVEEHVLSSFEAPNLNNEVIEDDDL
GDLINERDSRKRNNRRFDESNINNNNETKRNKYSSPVSGFYDHDINGY*
[0501] APJ1 is encoded by a non-essential gene comprising an ORF
that is 1.587 kbp in size and is located on chromosome XIV. A
published nucleotide coding sequence of APJ1 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00069
ATGCAACAAAACACGTCTTTATATGACTCTTTGAACGTTACTGCCGCTGCATCCACATCT
GAGATTAAGAAAGCTTACAGGAACGCTGCATTAAAATATCATCCTGATAAAAACAATCAT
ACAGAAGAATCCAAGCGAAAGTTTCAAGAGATATGCCAGGCATACGAAATACTTAAAGAC
AATCGTTTAAGAGCTTTGTATGACCAGTACGGTACCACAGATGAAGTCCTGATTCAAGAG
CAGCAGGCGCAGGCGCAACGCCAACAAGCCGGGCCGTTCAGTTCATCCTCAAATTTCGAT
ACGGAAGCAATGTCATTCCCGGATCTATCTCCAGGTGATCTTTTCGCGCAGTTTTTTAAT
AGTTCTGCTACCCCCTCTTCTAATGGCTCCAAAAGCAGTTTTAATTTTAGCTTCAATAAT
AGCTCTACGCCGAGCTTCTCCTTTGTTAATGGCAGTGGCGTGAACAATCTGTACTCCTCG
TCAGCAAAATACAACTCCAACGATGAGGACCATCATTTGGATAGAGGCCCTGATATCAAA
CATAATCTAAAGTGCACATTGAAGGAACTCTACATGGGTAAGACTGCAAAGTTGGGTTTG
AATAGGACAAGGATTTGCAGTGTTTGTGATGGGCACGGTGGTCTAAAGAAATGCACTTGT
AAAACATGCAAAGGGCAAGGTATTCAAACCCAAACTAGGCGTATGGGACCTCTAGTACAA
AGTTGGTCTCAAACTTGTGCAGATTGCGGGGGTGCCGGGGTTTTTGTCAAAAATAAAGAT
ATTTGCCAACAGTGCCAAGGTCTTGGCTTCATTAAGGAGAGGAAGATTCTACAAGTCACC
GTTCAACCGGGATCGTGTCATAACCAACTTATAGTACTTACGGGCGAAGGTGACGAAGTT
ATTAGTACTAAGGGAGGCGGTCACGAAAAGGTAATACCTGGTGACGTCGTTATCACCATT
TTACGTTTAAAAGATCCGAATTTCCAGGTTATCAACTACTCCAATTTGATATGTAAGAAG
TGCAAAATCGACTTCATGACCAGTTTATGTGGAGGCGTAGTTTATATTGAAGGGCACCCT
AGCGGTAAGTTGATCAAACTTGATATTATACCTGGCGAGATACTGAAGCCTGGTTGTTTC
AAGACTGTTGAGGACATGGGGATGCCCAAGTTTATCAACGGTGTTCGGAGCGGTTTCGGT
CATCTATATGTCAAATTCGATGTGACGTATCCAGAGAGACTGGAACCTGAAAATGCTAAG
AAAATACAAAATATTCTGGCTAATGATAAATACATTAAAGCAGAACGTTCCACCATGGAA
ACCGCAGATTCAGACTGCTATTGCGATTTGGAGAAGTCATATGACAGTGTGGAAGAGCAT
GTGTTAAGTAGCTTTGAGGCCCCTAATTTAAACAATGAAGTTATTGAAGACGACGACCTT
GGTGATTTGATTAATGAAAGAGATTCTCGGAAAAGGAACAACCGTCGATTCGACGAAAGT
AATATTAATAATAATAATGAAACGAAACGAAATAAATATTCTTCACCGGTAAGCGGTTTT
TATGACCATGATATTAATGGATATTGA
[0502] Further information on APJ1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000005021
[0503] It will be appreciated that, by "APJ1", we include fragments
or variants thereof having equivalent APJ1-like activity.
[0504] SIS1 is another S. cerevisiae helper protein of interest for
the present invention. It is a type II HSP40 co-chaperone that
interacts with the HSP70 protein Ssa1p; not functionally redundant
with Ydj1p due to due to substrate specificity; shares similarity
with bacterial DnaJ proteins. A published protein sequence for the
protein Sis1p is as follows:
TABLE-US-00070 MVKETKLYDLLGVSPSANEQELKKGYRKAALKYHPDKPTGDTEKFKEISE
AFEILNDPQKREIYDQYGLEAARSGGPSFGPGGPGGAGGAGGFPGGAGGF
SGGHAFSNEDAFNIFSQFFGGSSPFGGADDSGFSFSSYPSGGGAGMGGMP
GGMGGMHGGMGGMPGGFRSASSSPTYPEEETVQVNLPVSLEDLFVGKKKS
FKIGRKGPHGASEKTQIDIQLKPGWKAGTKITYKNQGDYNPQTGRRKTLQ
FVIQEKSHPNFKRDGDDLIYTLPLSFKESLLGFSKTIQTIDGRTLPLSRV
QPVQPSQTSTYPGQGMPTPKNPSQRGNLIVKYKVDYPISLNDAQKRAI DENF*
[0505] SIS1 is encoded by a non-essential gene comprising an ORF
that is 1.059 kbp in size and is located on chromosome XIV. A
published nucleotide coding sequence of SIS1 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00071
ATGGTCAAGGAGACAAAACTTTATGATTTACTTGGAGTATCTCCAAGTGCTAATGAGCAA
GAACTGAAAAAGGGTTATAGAAAAGCAGCTCTAAAATATCATCCAGATAAGCCAACAGGT
GACACAGAAAAGTTTAAGGAGATATCAGAGGCCTTTGAAATTTTAAATGATCCTCAAAAA
AGGGAAATATATGATCAATACGGTCTCGAGGCTGCTAGATCTGGTGGTCCAAGCTTTGGT
CCTGGTGGTCCTGGCGGTGCTGGAGGTGCTGGAGGCTTCCCTGGCGGTGCGGGCGGATTC
TCCGGAGGACATGCGTTCAGTAATGAGGATGCTTTCAATATTTTTTCACAATTCTTTGGC
GGCAGTTCCCCATTCGGTGGTGCTGATGACAGTGGCTTCAGTTTCTCTAGTTATCCATCT
GGCGGCGGTGCTGGTATGGGAGGTATGCCTGGAGGAATGGGAGGAATGCATGGCGGCATG
GGAGGTATGCCTGGCGGCTTTAGATCAGCATCAAGCTCTCCCACGTATCCAGAGGAAGAA
ACAGTTCAAGTTAATTTACCAGTTAGTCTAGAAGATTTGTTTGTTGGTAAAAAGAAGTCA
TTTAAAATTGGAAGAAAGGGCCCACATGGGGCCTCTGAAAAGACACAAATTGACATTCAA
TTAAAACCGGGTTGGAAAGCTGGTACCAAAATAACATACAAGAACCAGGGTGATTACAAT
CCTCAAACGGGCCGTAGAAAGACTTTGCAGTTTGTCATCCAGGAAAAGAGCCATCCAAAC
TTTAAAAGAGACGGTGATGACCTAATTTACACTCTGCCACTATCTTTCAAGGAATCATTG
TTAGGTTTTTCAAAAACTATCCAAACAATTGATGGCAGAACCTTACCTTTGTCGAGAGTA
CAGCCTGTCCAACCCTCACAAACTTCTACTTATCCTGGTCAAGGTATGCCAACTCCAAAG
AACCCATCTCAGAGAGGTAATTTGATTGTAAAATATAAAGTGGACTATCCAATATCACTA
AACGACGCTCAAAAACGTGCTATAGATGAAAATTTTTAA
[0506] Further information on SIS1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000004952
[0507] It will be appreciated that, by "SIS1", we include fragments
or variants thereof having equivalent SIS1-like activity.
[0508] DJP1 is another S. cerevisiae helper protein of interest for
the present invention. It is also known as ICS1 and PAS22. It is a
J-domain-containing protein, required for peroxisomal protein
import and involved in peroxisome assembly, homologous to E. coli
DnaJ and is located in the cytoplasm. A published protein sequence
for the protein Djp1p is as follows:
TABLE-US-00072
MVVDTEYYDLLGVSTTASSIEIKKAYRKKSIQEHPDKNPNDPTATERFQAISEAYQVLGD
DDLRAKYDKYGRKEAIPQGGFEDAAEQFSVIFGGDAFASYIGELMLLKNLQKTEELNAED
EAEKEKENVETMEESPADGKTNGTTNAVDAALGNTNEKDDKNKARTTSGNLTVHDGNKKN
EQVGAEAKKKKTKLEQFEEEQEVEKQKRVDQLSKTLIERLSILTESVYDDACKDSFKKKF
EEEANLLKMESFGLDILHTIGDVYYEKAEIFLASQNLFGMGGIFHSMKAKGGVFMDTLRT
VSAAIDAQNTMKELEKMKEASTNNEPLFDKDGNEQIKPTTEELAQQEQLLMGKVLSAAWH
GSKYEITSTLRGVCKKVLEDDSVSKKTLIRRAEAMKLLGEVFKKTFRTKVEQEEAQIFEE
LVAEATKKKRHT*
[0509] DJP1 is encoded by a non-essential gene comprising an ORF
that is 1.299 kbp in size and is located on chromosome IX. A
published nucleotide coding sequence of DJP1 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00073
ATGGTTGTTGATACTGAGTATTACGATTTGTTAGGTGTGTCTACCACTGCATCTTCCATT
GAAATAAAAAAGGCCTATAGAAAGAAATCTATTCAAGAGCATCCTGATAAGAATCCCAAT
GACCCCACGGCTACCGAAAGGTTTCAAGCAATATCCGAAGCTTATCAAGTTTTAGGTGAC
GATGATCTTCGCGCAAAGTATGATAAGTATGGAAGAAAAGAAGCTATTCCTCAGGGCGGC
TTTGAAGATGCAGCTGAACAGTTCTCTGTCATCTTTGGTGGAGATGCGTTTGCCTCATAT
ATTGGCGAACTGATGCTATTAAAGAACCTACAGAAAACTGAGGAGCTAAATGCTGAAGAC
GAAGCTGAAAAGGAGAAGGAGAATGTGGAAACAATGGAAGAATCACCTGCAGACGGTAAG
ACGAATGGCACCACTAACGCTGTTGATGCAGCATTGGGCAATACTAACGAAAAAGATGAC
AAAAATAAGGCGAGGACAACTTCTGGTAATTTAACTGTACACGATGGAAACAAGAAAAAT
GAGCAGGTAGGAGCAGAAGCTAAGAAGAAGAAGACAAAATTAGAGCAGTTTGAGGAAGAA
CAAGAGGTAGAAAAGCAAAAAAGAGTAGACCAATTAAGCAAAACATTGATTGAAAGATTA
TCGATATTAACAGAAAGTGTCTATGATGATGCATGTAAAGATTCCTTTAAAAAAAAGTTC
GAAGAGGAAGCCAATCTTTTAAAGATGGAATCATTTGGTCTGGACATATTACACACAATA
GGCGACGTTTACTACGAAAAAGCTGAAATTTTTCTTGCATCCCAGAACCTGTTCGGAATG
GGTGGTATATTTCATTCTATGAAGGCTAAAGGGGGAGTATTTATGGATACACTAAGAACT
GTTTCGGCAGCCATAGACGCTCAGAATACTATGAAGGAGCTTGAAAAAATGAAAGAAGCT
AGCACGAATAATGAGCCTTTGTTTGACAAAGACGGAAATGAGCAAATTAAGCCAACCACT
GAGGAACTGGCGCAGCAAGAGCAGCTATTGATGGGCAAAGTATTGTCGGCTGCTTGGCAT
GGTTCTAAATATGAAATAACATCCACTTTACGTGGCGTTTGTAAAAAAGTACTAGAAGAT
GACTCGGTAAGTAAGAAAACGCTTATCAGAAGAGCTGAAGCAATGAAACTATTGGGTGAA
GTCTTTAAGAAAACTTTCAGAACCAAAGTCGAACAAGAAGAGGCACAGATCTTTGAAGAA
CTTGTAGCAGAAGCTACAAAAAAGAAGAGACATACATGA
[0510] Further information on DJP1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000001443
[0511] It will be appreciated that, by "DJP1", we include fragments
or variants thereof having equivalent DJP1-like activity.
[0512] ZUO1 is another S. cerevisiae helper protein of interest for
the present invention. It is a cytosolic ribosome-associated
chaperone that acts, together with Ssz1p and the Ssb proteins, as a
chaperone for nascent polypeptide chains; contains a DnaJ domain
and functions as a J-protein partner for Ssb1p and Ssb2p. A
published protein sequence for the protein Zuo1p is as follows:
TABLE-US-00074
MFSLPTLTSDITVEVNSSATKTPFVRRPVEPVGKFFLQHAQRTLRNHTWSEFERIEAEKN
VKTVDESNVDPDELLFDTELADEDLLTHDARDWKTADLYAAMGLSKLRFRATESQIIKAH
RKQVVKYHPDKQSAAGGSLDQDGFFKIIQKAFETLTDSNKRAQYDSCDFVADVPPPKKGT
DYDFYEAWGPVFEAEARFSKKTPIPSLGNKDSSKKEVEQFYAFWHRFDSWRTFEFLDEDV
PDDSSNRDHKRYIERKNKAARDKKKTADNARLVKLVERAVSEDPRIKMFKEEEKKEKERR
KWEREAGARAEAEAKAKAEAEAKAKAESEAKANASAKADKKKAKEAAKAAKKKNKRAIRN
SAKEADYFGDADKATTIDEQVGLIVDSLNDEELVSTADKIKANAAGAKEVLKESAKTIVD
SGKLPSSLLSYFV*
[0513] ZUO1 is encoded by a non-essential gene comprising an ORF
that is 1.302 kbp in size and is located on chromosome VII. A
published nucleotide coding sequence of ZUO1 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00075
ATGTTTTCTTTACCTACCCTAACCTCAGACATCACTGTTGAAGTCAACAGTTCCGCTACC
AAAACCCCATTCGTCCGTCGTCCGGTCGAACCGGTTGGTAAGTTCTTTTTGCAACATGCT
CAAAGAACTTTGAGAAACCACACCTGGTCTGAATTTGAAAGAATTGAAGCTGAAAAGAAC
GTCAAAACCGTTGATGAATCCAATGTCGACCCAGATGAGTTGTTATTCGACACTGAATTG
GCCGATGAAGATTTACTGACTCATGATGCTAGAGACTGGAAAACTGCCGATTTGTATGCT
GCTATGGGTTTGTCTAAGTTGCGTTTCAGAGCTACTGAAAGTCAAATCATCAAGGCTCAC
AGAAAACAAGTTGTCAAGTACCATCCAGACAAGCAATCTGCTGCTGGTGGTAGTTTGGAC
CAAGATGGCTTTTTCAAGATTATTCAAAAGGCCTTTGAAACTTTGACTGATTCCAACAAG
AGAGCTCAGTACGACTCATGTGATTTTGTTGCCGATGTTCCTCCTCCAAAGAAGGGTACC
GATTATGACTTTTATGAAGCTTGGGGCCCCGTTTTCGAAGCTGAAGCTCGTTTTTCTAAG
AAGACTCCTATTCCTTCTCTAGGTAACAAAGATTCTTCCAAGAAGGAAGTTGAACAATTC
TATGCTTTCTGGCACAGATTTGACTCCTGGAGAACCTTTGAGTTCTTGGACGAAGATGTC
CCAGATGACTCTTCTAACAGAGACCACAAGCGTTACATTGAAAGAAAGAACAAGGCCGCA
AGAGACAAGAAGAAGACTGCTGATAACGCTAGATTGGTCAAACTTGTTGAAAGAGCTGTC
AGTGAAGATCCCCGTATCAAAATGTTCAAAGAAGAAGAGAAGAAGGAAAAGGAAAGAAGA
AAATGGGAAAGAGAAGCCGGTGCCAGAGCTGAAGCTGAAGCTAAGGCCAAGGCCGAAGCT
GAAGCGAAGGCTAAAGCTGAATCTGAAGCCAAGGCTAACGCCTCCGCAAAAGCTGACAAA
AAGAAGGCTAAGGAAGCTGCTAAGGCCGCCAAGAAAAAGAACAAGAGAGCCATCCGTAAC
TCTGCTAAGGAAGCTGACTACTTTGGTGATGCTGACAAGGCCACCACGATTGACGAACAA
GTTGGTTTGATCGTTGACAGTTTGAATGACGAAGAGTTAGTGTCCACCGCCGATAAGATC
AAGGCCAATGCTGCTGGTGCCAAGGAAGTTTTGAAGGAATCTGCAAAGACTATTGTCGAT
TCTGGCAAACTACCATCCAGCTTGTTGTCCTACTTCGTGTGA
[0514] Further information on ZUO1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000003517
[0515] It will be appreciated that, by "ZUO1", we include fragments
or variants thereof having equivalent ZUO1-like activity.
[0516] SWA2 is another S. cerevisiae helper protein of interest for
the present invention. It is also known as AUX1 and BUD24. It is an
auxilin-like protein involved in vesicular transport;
clathrin-binding protein required for uncoating of clathrin-coated
vesicles. A published protein sequence for the protein Swa2p is as
follows:
TABLE-US-00076
MSDPFAHLLTSLKNKDSASASKETTPQSSNSPSITGSAVADVARTDKSPNDSLHSISAPP
LIPSPKVDFSAPPLVPTNSTTKSNTANNTPPSALANTDDDFNQLFGMGTVTTTDTIQKPD
EDYYGSKEDHLYNGDDALVDEVKDMEIARLMSLGLSIEEATEFYENDVTYERYLEILKSK
QKERNDLAIRKKESGIKMEKSGLSNIVGTDSNNLFSMATDFFNKGKKLVDQWTSFPPEAN
DRLNNYSKTHDKVEDYDLPQVNDSPNRILFEDNEVVENLPPADNPDQDLLTDFETKIDIT
KRTAPDVSHSSSPTSGILIEENSRRNEPLIEDSLLDFSEGNLTNSKSNEDSTLFNENSNT
DSTIPISDIELSGYNEFKAKGTSLFKNGDYINSLQEYEKSLNTLPLNHPLRIIALSNIIA
SQLKIGEYSKSIENSSMALELFPSSKAKWKNKISNSDPERSFNDIWPKIMIRRAESFEHL
ESFKKALETYQELIKKNFFDDKIMQGKRRCQDFINPPPVKKSMPVKKKTTTTSPATKKQN
LTASSSNSPISVDSTSEIKKRELENAKLALYDKVFEKISSWKDGKDDDIRHLLANLSSLL
TWCNWKDVSMQDLVMPKRVKITYMKAVAKTHPDKIPESLSLENKMIAENIFSTLSIAWDK
FKLQNDIN*
[0517] SWA2 is encoded by a non-essential gene comprising an ORF
that is 2.007 kbp in size and is located on chromosome IV. A
published nucleotide coding sequence of SWA2 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00077
ATGTCAGATCCATTTGCACATTTACTGACTTCTTTGAAGAATAAGGACTCTGCATCTGCA
TCCAAGGAAACAACTCCTCAGAGCAGCAATTCGCCTTCCATTACTGGTTCCGCTGTTGCA
GATGTTGCAAGGACGGATAAAAGCCCCAATGATAGTCTGCATTCAATTTCAGCTCCTCCG
CTGATACCGTCACCGAAGGTAGATTTTTCTGCACCTCCTTTGGTCCCAACTAATAGCACC
ACTAAATCTAATACTGCCAACAACACACCTCCCTCGGCTCTTGCCAATACCGATGACGAC
TTCAATCAACTATTTGGTATGGGCACAGTAACTACAACGGATACGATCCAAAAACCGGAT
GAGGATTACTATGGAAGCAAGGAAGACCACCTTTACAATGGTGATGACGCCTTAGTTGAT
GAAGTTAAGGATATGGAAATAGCAAGATTGATGTCTCTAGGTTTATCAATTGAAGAAGCC
ACTGAGTTTTACGAAAATGACGTAACTTATGAAAGATATTTGGAGATTTTAAAGTCAAAG
CAAAAGGAGCGCAACGATCTAGCTATAAGAAAGAAAGAAAGTGGTATAAAAATGGAAAAG
TCAGGATTATCCAACATTGTTGGTACAGATAGCAATAATTTATTCAGCATGGCCACTGAT
TTTTTCAATAAGGGTAAGAAACTGGTAGACCAATGGACCTCCTTCCCACCTGAGGCAAAT
GATAGACTGAATAATTACTCAAAAACTCATGATAAGGTTGAGGATTATGATTTGCCTCAA
GTAAACGACTCACCCAATAGAATTTTGTTTGAAGATAATGAAGTCGTAGAGAACTTACCA
CCTGCCGATAATCCGGATCAAGATCTTTTAACTGATTTCGAAACAAAGATTGATATAACA
AAGAGGACAGCGCCTGATGTCTCCCACTCCTCCTCACCGACTTCTGGTATACTAATTGAA
GAAAATTCGCGAAGAAATGAGCCCCTGATAGAGGATAGTCTTCTCGACTTTTCAGAAGGA
AATCTCACCAATAGTAAAAGCAATGAAGATAGCACCCTCTTCAATGAAAACAGCAACACT
GACTCTACAATACCCATCTCAGATATTGAATTATCGGGGTATAACGAATTTAAGGCGAAA
GGTACTAGTTTGTTCAAGAACGGGGATTATATTAACTCATTACAAGAATATGAAAAGTCT
TTAAATACATTGCCTTTAAATCATCCATTGAGGATCATTGCATTATCAAACATTATTGCC
TCGCAACTGAAAATCGGTGAGTACTCTAAGTCCATAGAAAACTCCAGCATGGCTTTGGAA
TTATTCCCATCAAGCAAAGCTAAGTGGAAGAATAAAATCTCAAATAGTGACCCTGAAAGA
TCATTTAACGACATCTGGCCAAAGATTATGATTAGGCGTGCTGAGTCTTTTGAACATTTA
GAAAGTTTCAAAAAAGCACTAGAAACATACCAAGAGCTGATTAAGAAGAATTTTTTTGAT
GATAAAATCATGCAGGGAAAAAGAAGATGCCAAGACTTTATTAATCCTCCCCCTGTTAAA
AAATCCATGCCCGTTAAGAAGAAGACAACGACAACCTCGCCTGCAACAAAAAAACAGAAC
TTAACCGCTTCTTCTTCAAATTCTCCAATTTCTGTTGATAGCACTTCAGAAATAAAAAAA
CGGGAGCTAGAAAACGCTAAACTGGCGCTATATGATAAAGTATTTGAGAAAATTAGCTCC
TGGAAGGATGGCAAAGACGATGACATTCGTCATCTGTTAGCAAATTTATCCAGCTTACTA
ACATGGTGCAATTGGAAGGATGTCTCTATGCAAGATTTGGTTATGCCTAAGAGGGTCAAA
ATTACATACATGAAAGCTGTAGCCAAGACACATCCTGATAAGATACCAGAGTCCTTGTCC
CTGGAAAATAAGATGATTGCAGAGAATATTTTCAGTACTTTAAGTATTGCTTGGGATAAG
TTCAAACTGCAGAATGACATTAACTGA
[0518] Further information on SWA2 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000002728
[0519] It will be appreciated that, by "SWA2", we include fragments
or variants thereof having equivalent SWA2-like activity.
[0520] JJJ1 is another S. cerevisiae helper protein of interest for
the present invention. It contains a 70 amino acid J-domain, may
function as a co-chaperone to recruit Hsp70-like activity to
specific sites; mutation of it causes defects in fluid-phase
endocytosis. A published protein sequence for the protein Jjj1p is
as follows:
TABLE-US-00078
MKTCYYELLGVETHASDLELKKAYRKKALQYHPDKNPDNVEEATQKFAVIRAAYEVLSDP
QERAWYDSHKEQILNDTPPSTDDYYDYEVDATVTGVTTDELLLFFNSALYTKIDNSAAGI
YQIAGKIFAKLAKDEILSGKRLGKFSEYQDDVFEQDINSIGYLKACDNFINKTDKLLYPL
FGYSPTDYEYLKHFYKTWSAFNTLKSFSWKDEYMYSKNYDRRTKREVNRRNEKARQQARN
EYNKTVKRFVVFIKKLDKRMKEGAKIAEEQRKLKEQQRKNELNNRRKFGNDNNDEEKFHL
QSWQTVKEENWDELEKVYDNFGEFENSKNDKEGEVLIYECFICNKTFKSEKQLKNHINTK
LHKKNMEEIRKEMEEENITLGLDNLSDLEKFDSADESVKEKEDIDLQALQAELAEIERKL
AESSSEDESEDDNLNIEMDIEVEDVSSDENVHVNTKNKKKRKKKKKAKVDTETEESESFD
DTKDKRSNELDDLLASLGDKGLQTDDDEDWSTKAKKKKGKQPKKNSKSTKSTPSLSTLPS
SMSPTSAIEVCTTCGESFDSRNKLFNHVKIAGHAAVKNVVKRKKVKTKRI*
[0521] JJJ1 is encoded by a non-essential gene comprising an ORF
that is 1.773 kbp in size and is located on chromosome XIV. A
published nucleotide coding sequence of JJJ1 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00079
ATGAAGACCTGCTACTATGAGCTTTTAGGGGTCGAAACGCATGCTTCTGATCTTGAGTTA
AAAAAAGCTTACCGTAAAAAGGCCCTACAATATCACCCAGATAAAAACCCAGATAATGTT
GAAGAAGCCACACAAAAATTTGCTGTGATTCGAGCCGCTTATGAAGTACTGTCTGACCCC
CAGGAAAGAGCATGGTATGACTCACATAAGGAACAAATTTTAAATGATACTCCACCAAGC
ACTGATGATTACTATGATTATGAGGTAGACGCTACAGTCACAGGTGTCACAACTGATGAA
TTACTCTTATTTTTTAACTCTGCTCTTTATACTAAAATAGACAACTCAGCTGCTGGGATA
TATCAAATTGCAGGAAAAATATTTGCCAAGTTAGCTAAAGATGAGATTTTAAGTGGTAAG
CGACTGGGGAAATTTTCCGAGTATCAAGATGATGTATTCGAACAGGATATTAATAGTATT
GGCTATTTGAAAGCCTGCGATAACTTTATTAACAAGACGGATAAACTTTTATATCCTTTA
TTTGGATATTCGCCAACGGATTATGAATATTTGAAACATTTCTATAAGACTTGGTCAGCG
TTCAATACCTTGAAAAGTTTTAGCTGGAAAGACGAGTACATGTACTCTAAAAACTATGAC
AGAAGAACCAAGAGGGAAGTTAATAGAAGAAATGAGAAGGCTAGGCAACAAGCTCGAAAT
GAATACAACAAAACCGTGAAAAGGTTTGTAGTTTTCATAAAAAAGCTCGATAAAAGAATG
AAAGAAGGTGCAAAAATTGCAGAAGAACAGCGTAAACTAAAAGAACAACAGAGGAAAAAT
GAGTTAAATAACAGAAGAAAGTTTGGGAACGACAACAATGACGAAGAAAAATTTCATTTA
CAAAGCTGGCAAACGGTAAAAGAAGAAAACTGGGATGAACTGGAAAAGGTATATGATAAT
TTTGGAGAATTCGAAAATTCTAAGAATGATAAGGAAGGTGAAGTATTGATTTACGAGTGT
TTTATCTGCAACAAGACATTTAAGTCGGAAAAGCAATTGAAAAACCACATAAACACTAAA
CTGCATAAGAAAAATATGGAAGAGATACGGAAAGAAATGGAAGAGGAAAACATAACGCTT
GGGTTGGATAATCTCTCCGATCTCGAGAAATTTGATTCAGCAGATGAAAGTGTTAAAGAA
AAAGAAGATATTGATCTGCAAGCATTGCAAGCTGAACTCGCTGAAATTGAAAGAAAACTG
GCAGAATCGTCTTCTGAAGACGAAAGTGAAGATGACAATCTCAACATAGAAATGGATATA
GAGGTAGAAGACGTCAGTTCGGATGAAAATGTACATGTGAATACGAAGAATAAAAAGAAA
AGAAAAAAGAAAAAAAAAGCAAAGGTTGACACAGAAACAGAGGAATCTGAATCGTTCGAT
GATACTAAAGACAAACGGAGTAATGAGTTGGATGATCTTTTGGCATCACTAGGAGACAAG
GGCTTACAAACGGATGACGATGAAGATTGGTCTACTAAAGCGAAAAAGAAAAAGGGCAAA
CAACCTAAAAAGAATTCTAAATCCACAAAAAGCACTCCGTCCTTGTCGACTCTACCGTCC
TCTATGTCTCCAACCTCCGCGATCGAGGTGTGCACTACATGCGGAGAATCATTTGATAGT
CGAAATAAGCTATTCAACCACGTGAAGATAGCAGGGCATGCGGCAGTGAAAAACGTAGTG
AAAAGAAAGAAAGTCAAGACCAAAAGAATATAG
[0522] Further information on JJJ1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000005171
[0523] It will be appreciated that, by "JJJ1", we include fragments
or variants thereof having equivalent JJJ1-like activity.
[0524] JJJ2 is another S. cerevisiae helper protein of interest for
the present invention. It is one of several homologs of the
bacterial chaperone DnaJ, and is located in the cytoplasm. A
published protein sequence for the protein Jjj2p is as follows:
TABLE-US-00080
MSQVIEPQLDRTTYYSILGLTSNATSSEVHKSYLKLARLLHPDKTKSDKSEELFKAVVHA
HSILTDEDQKLRYDRDLKIKGLHTYQPKKNCHIFKTKAKESQGASPTLGQSEAYHRQNKP
YEQQPYGFGVGKKMTSSSKSKVPIFKSFNLKSYQRNHYYSSKKERKHGSPDIDSLFHETN
GASKVRMTDAGKMDTNSQFQEIWEILGKNAYTHKSYSEDPNSCLGSALSDHEEEEEAGKQ
QQQQQQQQQQQQHYGMTSKSSSPDEEKKNNKEPKRESRVSPEENGEEETGHKQFKLPKTS
TFSSGSHDSNLQSPFYNHEYRHYARSKFECKNQFRKSVSPIKEIPATTSANEGWNILRDI
IEKLNISNVDDRNKDLLFRRDEIGDKNHSDSIDIENLSIKEPKGMKRRKKDDISLEELFQ
SLPREKDYFMMDAINDSLESINLFKKPKTTQSHEQGGTFAQAESNRAKFKPLLEQCGITP
EILDLEIPEIPEFDAVADLETLKLNVQLFNNQCNKLKETIHQVSLQRLRADTQFSDMLTQ
KQSIMVWKTYLEFDKSLMDKLNILQERQMQVIKIFSERCDGKV*
[0525] JJJ2 is encoded by a non-essential gene comprising an ORF
that is 1.752 kbp in size and is located on chromosome 10. A
published nucleotide coding sequence of JJJ2 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00081
ATGTCACAGGTAATAGAACCACAATTAGATAGAACAACCTATTATTCCATATTAGGCTTG
ACATCAAATGCGACTTCCTCCGAAGTACATAAATCATATCTAAAACTGGCCAGATTACTT
CACCCAGATAAAACAAAATCTGATAAGTCTGAGGAATTATTCAAAGCTGTGGTGCATGCA
CATTCAATTTTAACTGATGAAGATCAAAAACTTCGATATGATCGAGATTTGAAAATCAAA
GGTTTACACACTTACCAGCCGAAGAAAAACTGTCATATTTTCAAGACCAAGGCAAAGGAA
TCACAAGGGGCTAGTCCCACACTTGGTCAATCAGAAGCTTATCATAGGCAAAATAAACCT
TATGAGCAACAGCCCTACGGTTTCGGTGTAGGCAAAAAAATGACCTCAAGCTCTAAGAGT
AAGGTTCCGATATTCAAGTCCTTCAATTTAAAAAGCTACCAACGAAACCACTATTATTCA
TCCAAAAAGGAAAGGAAACATGGAAGTCCTGATATTGATTCTTTGTTCCATGAAACCAAT
GGAGCCTCAAAAGTAAGAATGACTGATGCCGGTAAAATGGATACGAACTCTCAGTTCCAA
GAAATATGGGAAATATTGGGTAAAAATGCGTACACACATAAATCTTACTCTGAAGATCCA
AATTCATGTTTGGGATCAGCACTAAGCGATCATGAAGAAGAAGAAGAAGCAGGAAAACAA
CAACAGCAACAGCAGCAACAACAGCAACAGCAGCAACATTATGGAATGACGTCGAAGTCT
AGCAGTCCTGATGAAGAAAAAAAAAATAATAAAGAACCGAAAAGGGAAAGCAGAGTCTCT
CCAGAGGAAAATGGCGAAGAAGAAACGGGACACAAACAATTTAAATTGCCCAAGACCAGT
ACTTTTTCTAGTGGATCCCATGATTCAAATTTGCAATCTCCTTTTTACAATCATGAGTAT
CGACATTACGCAAGAAGTAAATTCGAATGCAAGAATCAGTTTAGAAAGTCAGTTTCTCCC
ATTAAAGAGATACCTGCAACAACTAGTGCCAATGAAGGATGGAACATTTTGAGAGACATT
ATTGAAAAACTCAATATAAGCAATGTAGACGATCGAAATAAAGACTTGCTGTTTCGTCGG
GATGAAATAGGTGATAAAAATCACAGCGACTCAATCGACATAGAAAATTTATCTATCAAA
GAACCTAAAGGGATGAAAAGGAGAAAGAAAGATGATATATCTTTAGAAGAATTGTTCCAA
TCTTTACCAAGAGAAAAAGATTATTTTATGATGGATGCAATTAATGACTCGTTAGAATCA
ATCAATCTTTTTAAAAAGCCGAAGACCACTCAGAGTCACGAACAAGGTGGAACTTTTGCC
CAAGCAGAAAGTAATCGTGCAAAATTCAAACCGTTACTAGAACAGTGTGGAATTACACCC
GAGATCTTAGATTTGGAAATACCAGAGATTCCGGAATTTGATGCAGTGGCTGACCTTGAA
ACATTGAAGCTTAACGTGCAGCTGTTTAATAACCAATGTAACAAACTTAAAGAAACAATA
CATCAAGTATCATTACAGCGCCTGAGAGCAGATACGCAGTTCAGTGATATGTTAACCCAA
AAGCAAAGTATTATGGTTTGGAAAACATACCTAGAATTTGATAAAAGTTTAATGGACAAA
TTGAACATCTTACAAGAAAGACAGATGCAGGTCATTAAAATTTTTTCCGAAAGATGTGAC
GGTAAAGTATAA
[0526] Further information on JJJ2 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000003698
[0527] It will be appreciated that, by "JJJ2", we include fragments
or variants thereof having equivalent JJJ2-like activity.
[0528] JJJ3 is another S. cerevisiae helper protein of interest for
the present invention and is also known as DPH4. It is one of
several homologs of the bacterial chaperone DnaJ, and is located in
the cytoplasm. A published protein sequence for the protein Jjj3p
is as follows:
TABLE-US-00082 MSLVNSLTHYEILRIPSDATQDEIKKAYRNRLLNTHPDKLSKSIHDTVSN
VTINKIQDAYKILSNIKTRREYDRLILENYKRQGFHNCGDGLDEFSLDDF
SFDEDKLEFMMNCPRCQFVGGFHFSESLLDECIDNVDAMERSHSGYQ
LLTQCSACSLWLKVNFDIEEEQEGQ
[0529] JJJ3 is encoded by a non-essential gene comprising an ORF
that is 0.519 kbp in size and is located on chromosome X. A
published nucleotide coding sequence of JJJ3 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00083
ATGTCATTGGTGAATTCGTTAACACACTACGAAATTTTAAGAATTCCATCGGATGCAACA
CAAGATGAAATCAAAAAGGCATATAGGAATCGGTTACTAAATACGCACCCCGATAAACTT
TCTAAAAGCATACATGATACGGTTAGCAACGTCACAATCAATAAGATTCAAGATGCTTAT
AAAATACTATCGAATATAAAAACTCGTCGCGAATATGATAGGTTGATCCTTGAAAACTAT
AAACGCCAAGGATTTCATAATTGTGGTGATGGGCTGGATGAATTTTCCTTAGACGATTTC
TCATTTGATGAAGATAAGCTGGAGTTTATGATGAATTGTCCTCGCTGTCAATTTGTTGGT
GGTTTTCATTTTAGTGAGAGTTTGTTAGATGAATGCATTGATAATGTAGACGCTATGGAA
CGGAGTCATTCTGGTTATCAATTATTAACCCAATGTAGCGCATGCAGCTTATGGCTGAAG
GTTAATTTTGACATCGAGGAAGAGCAAGAAGGACAATAA
[0530] Further information on JJJ3 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000003858
[0531] It will be appreciated that, by "JJJ3", we include fragments
or variants thereof having equivalent JJJ3-like activity.
[0532] CAJ1 is another S. cerevisiae helper protein of interest for
the present invention. It is one of several homologs of the
bacterial chaperone DnaJ, and is located in the nucleus. A
published protein sequence for the protein Caj1p is as follows:
TABLE-US-00084 MVKETEYYDILGIKPEATPTEIKKAYRRKAMETHPDKHPDDPDAQAKFQA
VGEAYQVLSDPGLRSKYDQFGKEDAVPQQGFEDASEYFTAIFGGDGFKDW
IGEFSLFKELNEATEMFGKEDEEGTAATETEKADESTDGGMVKHDTNKAE
SLKKDKLSKEQREKLMEMEKKRREDMMKQVDELAEKLNEKISRYLIAVK
SNNLEEFTRKLDQEIEDLKLESFGLELLYLLARVYKTKANNFIMSKKTY
GISKIFTGTRDNARSVKSAYNLLSTGLEAQKAMEKMSEVNTDELDQYERA
KFESTMAGKALGVMWAMSKFELERKLKDVCNKILNDKKVPSKERIAKAK
AMLFIAHKFASARRSPEEAEEARVFEELILGEQEKEHKKHTVAR
[0533] CAJ1 is encoded by a non-essential gene comprising an ORF
that is 1.176 kbp in size and is located on chromosome V. A
published nucleotide coding sequence of CAJ1 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00085
ATGGTAAAGGAGACGGAGTATTATGATATTTTGGGCATCAAGCCTGAGGCCACGCCCACT
GAAATCAAAAAGGCCTATCGTAGAAAGGCTATGGAAACACATCCGGACAAGCATCCTGAT
GACCCAGATGCTCAAGCAAAGTTTCAAGCCGTAGGCGAGGCCTACCAAGTCTTAAGTGAT
CCAGGGCTTCGTTCCAAGTATGACCAGTTTGGTAAGGAGGATGCTGTTCCTCAGCAAGGA
TTTGAAGATGCTTCTGAATACTTTACAGCAATATTCGGTGGTGATGGCTTCAAAGATTGG
ATTGGAGAATTTTCTTTGTTCAAAGAGCTAAACGAGGCAACAGAAATGTTTGGAAAGGAA
GATGAGGAGGGTACAGCAGCCACTGAAACCGAAAAAGCAGATGAGAGCACTGATGGTGGA
ATGGTTAAGCATGACACTAATAAAGCTGAATCTTTGAAAAAAGATAAATTATCGAAGGAG
CAAAGAGAGAAGCTAATGGAAATGGAGAAAAAAAGACGGGAAGATATGATGAAACAAGTC
GACGAGTTGGCAGAAAAACTGAACGAAAAAATCTCTAGGTACTTAATTGCTGTGAAGTCC
AATAACTTGGAGGAATTTACGCGAAAACTAGATCAAGAAATCGAGGATTTGAAATTAGAA
AGTTTTGGTCTAGAGTTATTGTATTTATTGGCCAGGGTTTACAAGACAAAAGCGAATAAT
TTTATCATGTCCAAGAAGACTTACGGAATTTCTAAAATATTCACTGGTACACGCGACAAT
GCTAGATCTGTTAAATCAGCATACAATTTATTGTCTACAGGCTTAGAAGCTCAAAAAGCC
ATGGAAAAAATGAGTGAAGTCAATACTGACGAACTAGACCAATATGAACGTGCCAAATTT
GAGTCCACAATGGCTGGTAAGGCACTTGGTGTCATGTGGGCTATGTCGAAATTTGAACTG
GAAAGAAAACTAAAAGACGTTTGCAATAAGATTCTAAACGATAAAAAGGTCCCTTCCAAG
GAACGTATTGCAAAGGCAAAAGCAATGCTGTTTATTGCCCACAAGTTTGCCAGTGCTAGA
AGGTCACCAGAAGAAGCTGAAGAAGCTAGAGTTTTTGAAGAGCTAATCCTAGGTGAGCAG
GAGAAGGAACACAAAAAACATACTGTGGCCAGATAA
[0534] Further information on CAJ1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000000850
[0535] It will be appreciated that, by "CAJ1", we include fragments
or variants thereof having equivalent CAJ1-like activity.
[0536] CWC23 is another S. cerevisiae helper protein of interest
for the present invention. It is one of several homologs of the
bacterial chaperone DnaJ, and is located in the nucleus. A
published protein sequence for the protein Cwc23p is as
follows:
TABLE-US-00086 MPGHELEDVINQRLNLYDVLELPTPLDVHTIYDDLPQIKRKYRTLALKYH
PDKHPDNPSIIHKFHLLSTATNILTNADVRPHYDRWLIEFLRKTNDIERN
KLIQKLEESESSTIPTTTPHPDLLQIQRHGELLRKLKHFNLPYGDWKHLN
TQDQENASQHPYYDCSTLRIVLDNFLQSNNKSNCLSHLRNQVFITLSANE
IYDIYFSERNNYSKDDSIIIYTVFDTPITAQHVFRNWSSGNLIPTVKDIS
PLIPLHYYSDFNLETELNDDIARLVSNEPILLD
[0537] CWC23 is encoded by an essential gene comprising an ORF that
is 0.852 kbp in size and is located on chromosome VII. A published
nucleotide coding sequence of CWC23 is as follows, although it will
be appreciated that the sequence can be modified by degenerate
substitutions to obtain alternative nucleotide sequences which
encode an identical protein product:
TABLE-US-00087
ATGCCAGGACACGAATTGGAAGACGTAATAAATCAACGTTTGAACCTATATGATGTATTA
GAATTACCGACCCCCCTGGACGTCCATACCATCTACGATGATTTGCCCCAAATTAAACGC
AAATACAGGACCCTTGCCCTGAAGTATCATCCTGACAAACACCCGGACAATCCATCAATT
ATACACAAATTCCACTTATTATCGACCGCAACTAATATCCTCACCAATGCAGACGTGAGA
CCCCATTACGACCGCTGGTTAATTGAGTTCCTACGGAAAACAAACGACATTGAAAGAAAT
AAACTTATACAAAAGCTGGAAGAATCTGAATCGAGTACGATACCCACCACCACACCACAT
CCTGATTTATTGCAAATCCAACGCCACGGCGAGCTACTCAGGAAACTAAAACATTTCAAC
TTGCCCTATGGTGACTGGAAACATCTCAACACACAAGACCAAGAAAATGCTTCGCAACAT
CCGTATTACGATTGCTCTACTTTGAGAATTGTCCTTGACAACTTCCTGCAATCAAATAAT
AAATCAAACTGCTTATCTCATTTGCGCAATCAAGTATTCATCACGCTAAGTGCTAATGAA
ATCTACGACATCTACTTCTCTGAAAGAAACAACTACTCGAAGGATGATTCAATCATCATA
TATACTGTATTCGATACTCCCATCACAGCGCAGCACGTATTCCGAAACTGGTCAAGTGGG
AACCTCATACCCACGGTCAAGGATATTTCGCCCTTGATCCCGCTACATTACTACTCTGAT
TTTAATTTGGAGACGGAACTGAATGACGATATTGCAAGACTGGTCTCTAATGAACCTATC
CTACTCGACTAG
[0538] Further information on CWC23 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000003096
[0539] It will be appreciated that, by "CWC23", we include
fragments or variants thereof having equivalent CWC23-like
activity.
[0540] PAM18 is another S. cerevisiae helper protein of interest
for the present invention and is also known as TIM14. It is one of
several homologs of the bacterial chaperone DnaJ, and is located in
the mitochondria. A published protein sequence for the protein
Pam18p is as follows:
TABLE-US-00088 MSSQSNTGNSIEAPQLPIPGQTNGSANVTVDGAGVNVGIQNGSQGQKTGM
DLYFDQALNYMGEHPVITGFGAFLTLYFTAGAYKSISKGLNGGKSTTAFL
KGGFDPKMNSKEALQILNLTENTLTKKKLKEVHRKIMLANHPDKGGSPFL
ATKINEAKDFLEKRGISK
[0541] PAM18 is encoded by an essential gene comprising an ORF that
is 0.507 kbp in size and is located on chromosome XII. A published
nucleotide coding sequence of PAM18 is as follows, although it will
be appreciated that the sequence can be modified by degenerate
substitutions to obtain alternative nucleotide sequences which
encode an identical protein product:
TABLE-US-00089
ATGAGTTCTCAAAGTAATACTGGTAATTCTATTGAGGCACCACAACTACCCATTCCTGGT
CAAACTAATGGCTCTGCGAACGTTACTGTTGATGGAGCTGGTGTTAATGTCGGTATCCAG
AATGGTTCGCAGGGTCAAAAGACCGGAATGGACCTTTATTTTGATCAAGCTTTGAACTAC
ATGGGAGAACATCCTGTGATAACAGGTTTTGGGGCCTTTTTAACTTTATATTTTACAGCC
GGTGCATATAAATCAATATCGAAGGGACTTAACGGTGGAAAATCCACTACTGCCTTCTTG
AAAGGCGGATTTGACCCGAAAATGAATTCTAAAGAGGCTCTACAGATTTTGAATTTGACA
GAAAATACATTGACTAAAAAAAAGTTGAAAGAGGTTCATAGGAAAATTATGTTAGCTAAT
CATCCTGACAAAGGTGGTTCTCCATTTTTGGCCACTAAGATAAACGAAGCTAAGGACTTT
TTGGAAAAAAGGGGTATTAGCAAATAA
[0542] Further information on PAM18 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000003998
[0543] It will be appreciated that, by "PAM18", we include
fragments or variants thereof having equivalent PAM18-like
activity.
[0544] JAC1 is another S. cerevisiae helper protein of interest for
the present invention. It is one of several homologs of the
bacterial chaperone DnaJ, and is located in the mitochondria. A
published protein sequence for the protein Jac1p is as follows:
TABLE-US-00090 MLKYLVQRRFTSTFYELFPKTFPKKLPIWTIDQSRLRKEYRQLQAQHHPD
MAQQGSEQSSTLNQAYHTLKDPLRRSQYMLKLLRNIDLTQEQTSNEVTTS
DPQLLLKVLDIHDELSQMDDEAGVKLLEKQNKERIQDIEAQLGQCYNDKD
YAAAVKLTVELKYWYNLAKAFKDWAPGKQLEMNH
[0545] JAC1 is encoded by an essential gene comprising an ORF that
is 0.555 kbp in size and is located on chromosome VII. A published
nucleotide coding sequence of JAC1 is as follows, although it will
be appreciated that the sequence can be modified by degenerate
substitutions to obtain alternative nucleotide sequences which
encode an identical protein product:
TABLE-US-00091
ATGTTGAAATACTTGGTTCAACGAAGATTCACTTCTACATTTTACGAGCTGTTCCCAAAG
ACCTTCCCCAAAAAGCTACCCATTTGGACTATCGATCAATCCAGATTAAGGAAGGAGTAT
AGGCAATTACAAGCACAGCACCATCCAGACATGGCCCAACAAGGTAGTGAACAGTCATCA
ACTCTTAATCAAGCTTACCATACTCTCAAAGATCCCCTTAGAAGGTCACAATATATGCTA
AAACTCTTGCGCAATATCGATTTGACGCAAGAACAGACCTCAAATGAAGTAACTACCAGT
GATCCACAGTTACTATTGAAAGTTCTAGACATCCATGATGAATTATCCCAGATGGACGAC
GAAGCTGGTGTGAAGCTGCTTGAAAAGCAAAACAAGGAAAGAATTCAAGATATTGAAGCC
CAGTTGGGACAATGCTACAATGACAAGGATTACGCCGCCGCAGTGAAGTTGACCGTGGAG
CTAAAGTACTGGTACAACTTGGCCAAGGCATTCAAAGACTGGGCTCCAGGAAAACAATTG
GAAATGAATCACTAA
[0546] Further information on JAC1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S0000002986
[0547] It will be appreciated that, by "JAC1", we include fragments
or variants thereof having equivalent JAC1-like activity.
[0548] JID1 is another S. cerevisiae helper protein of interest for
the present invention. It is one of several homologs of the
bacterial chaperone DnaJ, and is located in the mitochondria. A
published protein sequence for the protein Jid1p is as follows:
TABLE-US-00092 MLHHKFVYPFLFKWHLSCVEKCPPQITFIAKYATANDKNGNRKLTIRDEQ
WPELADPTPYDIFGIPKAGSGNPKLDKKSLKKKYHRYVKLYHPDHSDNIQ
IFSSEKVTNSDSKSPLLLTSSEKLHRFKVISQAYDILCDPKKKIVYDTTR
QGWTTSYSPRSNVNTENYQYAGSYGYHSNAQYEYWNAGTWEDANSMKN
ERIQENINPWTVIGIICGLAICIEGTALLAKIQESLSKAEFTHDESGLHL
IQSYTNYGLDTDKFSRLRRFLWFRTWGLYKSKEDLDREAKINEEMIRKLK AAK
[0549] JID1 is encoded by a non-essential gene comprising an ORF
that is 0.906 kbp in size and is located on chromosome XVI. A
published nucleotide coding sequence of JID1 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00093
ATGCTACACCATAAGTTCGTATACCCATTTTTATTCAAGTGGCACTTATCATGTGTAGAA
AAGTGTCCCCCACAAATCACTTTTATAGCTAAGTATGCTACAGCGAACGATAAAAATGGC
AATAGAAAACTTACGATAAGGGATGAACAATGGCCTGAGTTGGCAGATCCAACTCCCTAT
GATATTTTTGGCATTCCAAAGGCCGGATCTGGAAATCCTAAACTGGACAAGAAGTCGTTA
AAAAAAAAATATCATCGTTATGTAAAATTGTACCACCCTGACCATTCCGATAACATTCAA
ATATTTAGCTCAGAAAAGGTTACCAACAGTGATAGTAAATCACCGCTGCTGCTAACATCA
AGCGAAAAACTACATAGATTTAAAGTCATCTCTCAAGCATATGATATTCTTTGTGACCCA
AAGAAAAAGATCGTATATGACACAACGAGGCAAGGCTGGACCACATCGTATTCACCACGT
TCTAACGTTAATACTGAAAATTACCAATATGCCGGCTCTTATGGCTACCACTCTAACGCG
CAGTATGAATACTGGAACGCTGGGACTTGGGAAGACGCAAATAGCATGAAAAACGAAAGA
ATTCAAGAAAACATCAACCCATGGACCGTTATTGGCATAATTTGTGGCCTAGCTATATGC
ATCGAAGGGACTGCGTTGTTAGCCAAAATCCAGGAGTCTCTGAGCAAGGCCGAATTTACT
CATGACGAAAGTGGATTACATTTGATTCAGTCATACACGAATTATGGTCTTGATACTGAC
AAATTTTCCAGATTGAGGCGGTTCTTATGGTTTAGAACTTGGGGACTTTACAAGTCGAAA
GAGGATTTAGATAGAGAAGCCAAGATCAATGAAGAAATGATACGCAAACTGAAAGCAGCT
AAATGA
[0550] Further information on JID1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000006265
[0551] It will be appreciated that, by "JID1", we include fragments
or variants thereof having equivalent JID1-like activity.
[0552] HLJ1 is another S. cerevisiae helper protein of interest for
the present invention. It is one of several homologs of the
bacterial chaperone DnaJ, and is located in the endoplasmic
reticulum membrane. A published protein sequence for the protein
Hlj1p is as follows:
TABLE-US-00094 MSFTEDQEKIALEILSKDKHEFYEILKVDRKATDSEIKKAYRKLAIKLHP
DKNSHPKAGEAFKVINRAFEVLSNEEKRSIYDRIGRDPDDRQMPSRGAAS
GFRGSAGGSPMGGGFEDMFFNSRFGGQRAGPPEDIFDFLFNAGGSPFGA
SPFGPSASTFSFGGPGGFRVYTNNRGGSPFMRQQPRSRQQQQQAEENA
VNSQLKNMLVLFIIFIVLPMIKDYLFS
[0553] HLJ1 is encoded by a non-essential gene comprising an ORF
that is 0.675 kbp in size and is located on chromosome XIII. A
published nucleotide coding sequence of HLJ1 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00095 ATGTCTTTCACTGAGGATCAAGAAAAAATCGCGCTAGAAATACTGTCAAA
AGACAAGCATGAGTTTTACGAAATTTTGAAGGTAGATAGGAAAGCCACAG
ATAGTGAGATCAAGAAGGCATACAGAAAACTAGCAATCAAATTGCATCCT
GATAAAAACTCTCATCCAAAAGCGGGAGAAGCTTTCAAAGTAATTAATAG
GGCATTTGAAGTACTAAGCAATGAGGAAAAGCGCAGTATTTATGACAGGA
TAGGTAGGGATCCTGACGATAGACAAATGCCATCCAGAGGTGCTGCTTCA
GGGTTCCGAGGAAGTGCAGGTGGGTCTCCAATGGGTGGCGGATTTGAAGA
CATGTTTTTCAATTCACGTTTCGGTGGTCAAAGAGCTGGACCACCAGAGG
ACATATTCGACTTTTTGTTCAACGCAGGCGGCAGCCCATTCGGCGCTTCA
CCATTTGGGCCTTCTGCTTCCACTTTTTCATTTGGAGGCCCCGGTGGTTT
CAGAGTTTATACTAATAATCGTGGTGGCTCACCGTTCATGCGTCAACAAC
CCCGCTCAAGACAGCAGCAACAACAAGCAGAAGAAAATGCAGTGAATTCG
CAATTAAAAAATATGCTCGTTCTTTTCATCATCTTTATTGTTCTTCCTAT
GATTAAAGATTACCTGTTTAGTTAA
[0554] Further information HLJ1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000004771
[0555] It will be appreciated that, by "HLJ1", we include fragments
or variants thereof having equivalent HLJ1-like activity.
[0556] ERJ5 is another S. cerevisiae helper protein of interest for
the present invention. It is one of several homologs of the
bacterial chaperone DnaJ, and is located in the endoplasmic
reticulum. A published protein sequence for the protein Erj5p is as
follows:
TABLE-US-00096 MNGYWKPALVVLGLVSLSYAFTTIETEIFQLQNEISTKYGPDMNFYKFLK
LPKLQNSSTKEITKNLRKLSKKYHPDKNPKYRKLYERLNLATQILSNSSN
RKIYDYYLQNGFPNYDFHKGGFYFSRMKPKTWFLLAFIWIVVNIGQYIIS
IIQYRSQRSRIENFISQCKQQDDTNGLGVKQLTFKQHEKDEGKSLVVRFS
DVYVVEPDGSETLISPDTLDKPSVKNCLFWRIPASVWNMTFGKSVGSAGK
EEIITDSKKYDGNQTKKGNKVKKGSAKKGQKKMELPNGKVIYSRK
[0557] ERJ5 is encoded by a non-essential gene comprising an ORF
that is 0.888 kbp in size and is located on chromosome VI. A
published nucleotide coding sequence of ERJ5 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00097 ATGAACGGTTACTGGAAACCTGCGTTGGTTGTCCTGGGATTGGTATCTCT
ATCATATGCTTTTACCACCATTGAAACAGAAATTTTCCAATTACAAAATG
AAATAAGTACGAAATATGGCCCAGATATGAACTTCTACAAGTTCTTGAAG
TTACCTAAACTGCAGAATTCTAGTACAAAGGAGATTACAAAAAACTTAAG
AAAGCTATCCAAGAAGTACCATCCGGATAAGAACCCTAAATACCGTAAAT
TGTATGAAAGGTTAAACCTCGCTACTCAAATTCTTTCAAACAGCTCTAAT
CGTAAGATTTATGATTATTATCTACAGAATGGCTTTCCAAACTATGATTT
CCATAAGGGTGGTTTTTATTTTTCCAGAATGAAGCCTAAGACTTGGTTCC
TGCTGGCCTTTATTTGGATAGTCGTTAATATTGGGCAGTATATCATTTCT
ATTATTCAATATCGTTCTCAAAGATCAAGAATTGAAAACTTCATCAGTCA
GTGTAAACAACAGGATGATACCAATGGACTAGGCGTAAAACAACTAACGT
TTAAACAACATGAAAAGGATGAGGGTAAAAGTTTGGTTGTAAGGTTTAGC
GATGTCTATGTTGTAGAGCCTGATGGAAGTGAAACACTAATTTCGCCAGA
TACCTTGGATAAACCTTCAGTAAAGAACTGTTTGTTTTGGAGAATACCTG
CTTCGGTTTGGAACATGACGTTTGGCAAATCTGTTGGTAGCGCAGGAAAA
GAAGAAATAATAACGGATAGTAAAAAGTATGATGGTAACCAAACAAAAAA
GGGGAACAAAGTAAAAAAGGGTTCTGCAAAGAAAGGCCAAAAGAAAATGG
AATTGCCTAACGGTAAAGTGATCTATTCACGTAAATGA
[0558] Further information ERJ5 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000001937
[0559] It will be appreciated that, by "ERJ5", we include fragments
or variants thereof having equivalent ERJ5-like activity.
[0560] MGE1 is another S. cerevisiae helper protein of interest for
the present invention and is also known as YGE1. It is one of
several homologs of the bacterial GrpE and is located in the
mitochondria. A published protein sequence for the protein Mge1p is
as follows:
TABLE-US-00098 MRAFSAATVRATTRKSFIPMAPRTPFVTPSFTKNVGSMRRMRFYSDEAKS
EESKENNEDLTEEQSEIKKLESQLSAKTKEASELKDRLLRSVADFRNLQQ
VTKKDIQKAKDFALQKFAKDLLESVDNFGHALNAFKEEDLQKSKEISDLY
TGVRMTRDVFENTLRKHGIEKLDPLGEPFDPNKHEATFELPQPDKEPGTV
FHVQQLGFTLNDRVIRPAKVGIVKGEEN
[0561] MGE1 is encoded by an essential gene comprising an ORF that
is 0.687 kbp in size and is located on chromosome XV. A published
nucleotide coding sequence of MGE1 is as follows, although it will
be appreciated that the sequence can be modified by degenerate
substitutions to obtain alternative nucleotide sequences which
encode an identical protein product:
TABLE-US-00099 ATGAGAGCTTTTTCAGCAGCCACCGTTAGGGCCACAACTAGGAAGTCGTT
CATCCCAATGGCACCAAGAACTCCTTTTGTGACTCCATCATTTACAAAGA
ATGTAGGCTCAATGAGAAGAATGAGATTTTATTCTGATGAAGCCAAAAGT
GAAGAATCCAAAGAAAACAATGAAGATTTGACTGAAGAGCAATCAGAAAT
CAAGAAATTAGAGAGCCAGTTAAGCGCGAAGACTAAAGAAGCTTCTGAAC
TCAAGGACAGATTATTAAGATCTGTGGCAGATTTCAGAAATTTACAACAA
GTCACAAAGAAGGATATTCAGAAAGCTAAGGACTTTGCTTTACAGAAGTT
TGCAAAGGATTTATTGGAATCTGTAGATAACTTTGGTCATGCTTTGAATG
CTTTTAAAGAGGAAGACTTACAAAAGTCCAAGGAAATTAGTGATTTGTAT
ACAGGGGTTAGAATGACAAGAGATGTTTTTGAAAACACCCTAAGAAAGCA
CGGTATTGAAAAATTAGACCCATTGGGAGAACCATTTGATCCAAATAAAC
ACGAAGCAACGTTCGAGTTGCCACAACCTGATAAGGAACCGGGTACTGTT
TTCCATGTACAACAATTAGGTTTCACCTTGAATGACAGAGTTATCAGACC
AGCAAAAGTCGGAATTGTTAAGGGCGAAGAGAACTAA
[0562] Further information MGE1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000005758
[0563] It will be appreciated that, by "MGE1", we include fragments
or variants thereof having equivalent MGE1-like activity.
[0564] FES1 is another S. cerevisiae helper protein of interest for
the present invention. It is one of several homologs of the
bacterial GrpE and is located in the cytoplasm. A published protein
sequence for the protein Fes1p is as follows:
TABLE-US-00100 MEKLLQWSIANSQGDKEAMARAGQPDPKLLQQLFGGGGPDDPTLMKESMA
VIMNPEVDLETKLVAFDNFEMLIENLDNANNIENLKLWEPLLDVLVQTKD
EELRAAALSIIGTAVQNNLDSQNNFMKYDNGLRSLIEIASDKTKPLDVRT
KAFYALSNLIRNHKDISEKFFKLNGLDCIAPVLSDNTAKPKLKMRAIALL
TAYLSSVKIDENIISVLRKDGVIESTIECLSDESNLNIIDRVLSFLSHLI
SSGIKFNEQELHKLNEGYKHIEPLKDRLNEDDYLAVKYVL
[0565] FES1 is encoded by a non-essential gene comprising an ORF
that is 0.873 kbp in size and is located on chromosome II. A
published nucleotide coding sequence of FES1 is as follows,
although it will be appreciated that the sequence can be modified
by degenerate substitutions to obtain alternative nucleotide
sequences which encode an identical protein product:
TABLE-US-00101 ATGGAAAAGCTATTACAGTGGTCTATTGCGAATTCTCAAGGGGACAAAGA
AGCTATGGCTAGGGCCGGCCAACCTGATCCTAAATTGCTACAGCAGTTAT
TCGGTGGTGGTGGTCCTGACGATCCAACCTTAATGAAAGAATCCATGGCT
GTTATTATGAATCCGGAGGTTGACTTAGAAACAAAACTCGTTGCATTTGA
CAACTTTGAAATGTTGATTGAGAACTTAGATAATGCTAATAATATCGAAA
ATTTAAAACTGTGGGAGCCATTGTTGGATGTTCTTGTTCAGACGAAGGAT
GAAGAACTACGTGCTGCTGCTTTATCCATTATTGGAACGGCTGTGCAAAA
CAACTTGGATTCGCAAAATAATTTCATGAAATACGACAATGGTCTGCGAA
GCCTTATCGAAATAGCTAGTGACAAGACAAAGCCACTCGACGTGAGAACA
AAAGCTTTTTACGCACTATCTAATCTAATAAGAAACCACAAAGATATCTC
AGAAAAGTTTTTCAAATTAAATGGGCTCGACTGCATAGCACCTGTATTAA
GTGATAACACCGCCAAACCAAAACTGAAAATGAGAGCCATTGCCTTATTG
ACCGCATATTTGTCATCTGTTAAGATTGATGAAAATATAATCAGTGTGCT
GAGAAAGGATGGAGTAATTGAAAGTACGATTGAGTGCTTGTCTGACGAGA
GTAACTTGAACATCATAGATAGAGTTCTGTCTTTTCTCTCTCACCTGATA
TCTTCCGGAATAAAATTTAATGAACAGGAATTGCACAAATTGAACGAAGG
TTACAAACATATCGAGCCTCTAAAGGACAGACTTAATGAAGACGATTATT
TAGCCGTAAAGTATGTATTATGA
[0566] Further information FES1 can be obtained from the URL
address
http://db.yeastgenome.org/cgi-bin/singlepageformat?sgdid=S000000305
[0567] It will be appreciated that, by "FES1", we include fragments
or variants thereof having equivalent FES1-like activity.
[0568] Variants and fragments of the above JEM1, LHS1, SCJ1, KAR2,
SIL1, FKB2, SSA1, SSA2, SSA3, SSA4, SSE1, SSE2, SSB1, SSB2, ECM10,
MDJ1, MDJ2, ERO1, ERV2, EUG1, MPD1, MPD2, EPS1, PDI1, DER1, DER3,
HRD3, UBC7, DOA4, HAC1, SEC63, YDJ1, XDJ1, APJ1, SIS1, DJP1, ZUO1,
SWA2, JJJ1, JJJ2, JJJ3, CAJ1, CWC23, PAM18, JAC1, JID1, HLJ1, ERJ5,
MGE1 and FES1 proteins and encoding polynucleotide sequences, and
variants of other naturally occurring JEM1, LHS1, SCJ1, KAR2, SIL1,
FKB2, SSA1, SSA2, SSA3, SSA4, SSE1, SSE2, SSB1, SSB2, ECM10, MDJ1,
MDJ2, ERO1, ERV2, EUG1, MPD1, MPD2, EPS1, PDI1, DER1, DER3, HRD3,
UBC7, DOA4, HAC1, SEC63, YDJ1, XDJ1, APJ1, SIS1, DJP1, ZUO1, SWA2,
JJJ1, JJJ2, JJJ3, CAJ1, CWC23, PAM18, JAC1, JID1, HLJ1, ERJ5, MGE1
and FES1 proteins and encoding polynucleotide sequences are also
included in the present invention.
[0569] A "variant", in the context of a JEM1, LHS1, SCJ1, KAR2,
SIL1, FKB2, SSA1, SSA2, SSA3, SSA4, SSE1, SSE2, SSB1, SSB2, ECM10,
MDJ1, MDJ2, ERV2, EUG1, MPD1, MPD2, EPS1, PDI1, DER1, DER3, HRD3,
UBC7, DOA4, HAC1, SEC63, YDJ1, XDJ1, APJ1, SIS1, DJP1, ZUO1, SWA2,
JJJ1, JJJ2, JJJ3, CAJ1, CWC23, PAM18, JAC1, JID1, HLJ1, ERJ5, MGE1
or FES1 protein, refers to a protein having a sequence as defined
above by the present application wherein at one or more positions
there have been amino acid insertions, deletions, or substitutions,
either conservative or non-conservative, provided that such changes
result in a protein whose basic properties, for example enzymatic
activity (type of and specific activity), thermostability, activity
in a certain pH-range (pH-stability) have not significantly been
changed. "Significantly" in this context means that one skilled in
the art would say that the properties of the variant may still be
different but would not be unobvious over the ones of the original
protein.
[0570] By "conservative substitutions" is intended combinations
such as Val, Ile, Leu, Ala, Met; Asp, Glu; Asn, Gln; Ser, Thr, Gly,
Ala; Lys, Arg, His; and Phe, Tyr, Trp. Preferred conservative
substitutions include Gly, Ala; Val, Ile, Leu; Asp, Glu; Asn, Gln;
Ser, Thr; Lys, Arg; and Phe, Tyr.
[0571] A "variant" typically has at least 25%, at least 50%, at
least 60% or at least 70%, preferably at least 80%, more preferably
at least 90%, even more preferably at least 95%, yet more
preferably at least 99%, most preferably at least 99.5% sequence
identity to the polypeptide from which it is derived.
[0572] The percent sequence identity between two polypeptides may
be determined using suitable computer programs, as discussed below.
Such variants may be natural or made using the methods of protein
engineering and site-directed mutagenesis as are well known in the
art.
[0573] A "fragment", in the context of JEM1, LHS1, SCJ1, KAR2,
SIL1, FKB2, SSA1, SSA2, SSA3, SSA4, SSE1, SSE2, SSB1, SSB2, ECM10,
MDJ1, MDJ2, ERO1, ERV2, EUG1, MPD1, MPD2, EPS1, PDI1, DER1, DER3,
HRD3, UBC7, DOA4, HAC1, SEC63, YDJ1, XDJ1, APJ1, SIS1, DJP1, ZUO1,
SWA2, JJJ1, JJJ2, JJJ3, CAJ1, CWC23, PAM18, JAC1, JID1, HLJ1, ERJ5,
MGE1 and FES1 proteins, refers to a protein wherein at one or more
positions there have been deletions. Thus the fragment may comprise
at most 5, 10, 20, 30, 40 or 50%, typically up to 60%, more
typically up to 70%, preferably up to 80%, more preferably up to
90%, even more preferably up to 95%, yet more preferably up to 99%
of the complete sequence of the full mature protein as defined
above. Particularly preferred fragments of a protein comprise one
or more whole domains of the desired protein.
[0574] A fragment or variant of a JEM1, LHS1, SCJ1, KAR2, SIL1,
FKB2, SSA1, SSA2, SSA3, SSA4, SSE1, SSE2, SSB1, SSB2, ECM10, MDJ1,
MDJ2, ERO1, ERV2, EUG1, MPD1, MPD2, EPS1, PDI1, DER1, DER3, HRD3,
UBC7, DOA4, HAC1, SEC63, YDJ1, XDJ1, APJ1, SIS1, DJP1, ZUO1, SWA2,
JJJ1, JJJ2, JJJ3, CAJ1, CWC23, PAM18, JAC1, JID1, HLJ1, ERJ5, MGE1
or FES1 protein may be a protein that, when expressed recombinantly
in a host cell, can complement the deletion of the same
endogenously encoded gene in the host cell, such as S. cerevisiae,
and may or may not, for example, be a naturally occurring homolog
of the protein upon which it is based, such as a homolog encoded by
another organism, such as another yeast or other fungi, or another
eukaryote such as a human or other vertebrate, or animal or by a
plant.
[0575] A fragment or a variant of a polynucleotide encoding a JEM1,
LHS1, SCJ1, KAR2, SIL1, FKB2, SSA1, SSA2, SSA3, SSA4, SSE1, SSE2,
SSB1, SSB2, ECM10, MDJ1, MDJ2, ERO1, ERV2, EUG1, MPD1, MPD2, EPS1,
PDI1, DER1, DER3, HRD3, UBC7, DOA4, HAC1, SEC63, YDJ1, XDJ1, APJ1,
SIS1, DJP1, ZUO1, SWA2, JJJ1, JJJ2, JJJ3, CAJ1, CWC23, PAM18, JAC1,
JID1, HLJ1, ERJ5, MGE1 or FES1 protein may be a polynucleotide that
comprises a sequence that encodes a fragment or variant of the
protein as defined above.
[0576] The present invention will now be exemplified with reference
to the following non-limiting examples and figures.
BRIEF DESCRIPTION OF FIGURES
[0577] FIGS. 1 to 9, 11 to 16, 21, 23-25 and 28 show various
plasmid maps as described in the following examples.
[0578] FIG. 10 shows analysis of HAC1 splicing at log phase by
qRT-PCR in the strain AH22 (ura3) [pAYE329]. Helper protein
overexpression plasmids are shown on the x-axis. Data are
normalised to ACT1 transcript levels and presented as fold changes
from AH22 (ura3) [pAYE329, YCplac33]. All values shown represent
duplicate analysis of mRNA levels from single experimental
cultures.
[0579] FIG. 17 shows SDS-PAGE gels for quantification of rHA
production in overexpression strains. Sample labels shown indicate
overexpression plasmids transformed into the strain AH22 (ura3)
[pAYE329]. Duplicate samples represent two independent shake flasks
from the same transformant.
[0580] FIG. 18 shows quantification of main rHA band in transformed
and control strains, by analysis of SDS-PAGE gel of FIG. 17 using
densitometry. Values are normalised (based on culture optical
density readings) to account for different growth rates observed
between strains.
[0581] FIG. 19 shows quantification of main rHA band in transformed
and control strains, by analysis of SDS-PAGE gel of FIG. 17 using
densitometry, expressed as a percentage of determined rHA
production by the negative control YCplac33. Values are normalised
(based on culture optical density readings) to account for
different growth rates observed between strains.
[0582] FIG. 20 shows quantification of rHA fragments relative to
total rHA, by analysis of SDS-PAGE gel of FIG. 17 using
densitometry, expressed as a percentage of detected rHA fragments
relative to total rHA levels detected (total rHA=full length
rHA+degradation products). Values are normalised (based on culture
optical density readings) to account for different growth rates
observed between strains.
[0583] FIG. 22 shows a comparison of recombinant transferrin titres
by rocket immunoelectrophoresis. A=Control Strain [pDB3213];
B=Control Strain (ura3). [pTPC17 pDB3213]. Duplicate 10 mL shake
flasks cultures were inoculated with yeast and incubated with
shaking at 200 rpm for 4-days at 30.degree. C. 5 .mu.L culture
supernatant loaded per well of a rocket immunoelectrophoresis gel.
Plasma Tf standards concentrations are in .mu.g/mL. 20 .mu.L goat
anti-Tf/50 mL agarose. Precipin was stained with Coomassie
blue.
[0584] FIG. 26 shows the effect of LHS1, JEM1 and SIL1
co-expression on rHA production, when rHA is fused to different
leader sequences. Two separate transformants for each strain were
inoculated into 50 mL shake flasks containing 10 mL BMMD and
incubated with shaking at 200 rpm for 4-days at 30.degree. C. 20
.mu.L of culture supernatant was loaded per well of a 4-12%
SDS-PAGE gel and run for 50 mins in MOPS buffer. Gel A shows the
results obtained with plasmid pDB2244, which encodes a
HSA/MF.alpha.-1 fusion leader sequence (A=AH22 (ura3) [pDB2244
YCplac33]; B=AH22 (ura3) [pDB2244 pTPC17]). Gel B shows the results
obtained with plasmid pDB2286, which encodes an invertase leader
sequence (C=AH22 (ura3) [pDB2286 YCplac33]; D=AH22 (ura3) [pDB2286
pTPC17]). Gel C shows the results obtained with plasmid pDB2287,
which encodes the MF.alpha.-1 leader sequence (E=AH22 (ura3)
[pDB2287 YCplac33]; F=AH22 (ura3) [pDB2287 pTPC17]).
[0585] FIG. 26, part D, shows densitometric quantification of rHA
secretion. Gels shown in FIG. 26 A-C were analysed by densitometry
and comparison to rHA standard curves. Data presented above
represents quantification of single rHA bands. For each strain two
transformants were analysed (samples A and B in FIG. 26D).
[0586] FIG. 27 shows the DNA sequence of the human GM-CSF cDNA with
an incorporated N-terminal Met codon.
[0587] FIG. 29 A shows an SDS-PAGE gel for quantification of GM-CSF
production. Lanes 2-5 show GM-CSF production in the control strain
(ura3) [pDB2109 YCplac33]. Lanes 6-9 show GM-CSF production in the
control strain (ura3) [pDB2109 pTPC17.
[0588] FIG. 29 B shows the results of densitometric analysis of the
SDS-PAGE gel shown in FIG. 29 A, as further given in Table 9,
below.
EXAMPLE 1
[0589] A strain of S. cerevisiae that possesses increased
production of a recombinant protein was produced by the following
methodology.
[0590] Strains. The S. cerevisiae strain used was a histidine
revertant of AH22 (cir.sup.o a leu2-3 leu2-112 his4 canR). AH22 is
further described in Mead at al, 1986, Mol. Gen. Genet., 205,
417-421. A polynucleotide encoding a recombinant heterologous
protein expression cassette was introduced by S. cerevisiae
transformation performed according to Ito, H., at al.
(Transformation of intact yeast cells treated with alkali cations.
J. Bacterial. 153, 163-168, (1983)).
[0591] Media. Yeast strains were grown in rich broth medium, YEP
(1% yeast extract 2% w/v Bactopeptone).
[0592] Protein assays. Yeast cells were grown in 10 ml cultures for
72 hours to a density of 5.times.10.sup.7 cells/mL at 30.degree. C.
in YEP 2% (w/v) sucrose. In order to analyse the soluble
heterologous protein fraction of yeast, cells were harvested by
centrifugation and disrupted in phosphate buffered saline by
vortexing with 40 mesh glass beads. The soluble fraction was
collected as the supernatant of a 10,000.times.g centrifugation.
The fraction was assayed for the presence of heterologous protein
by polyacrylamide gel electrophoresis and Western blot, using
appropriate commercially available antibodies.
[0593] Mutagenesis. Yeast cells to be mutated were grown in 100 ml
defined medium (0.65% (w/v) YNB; 2% (w/v) sucrose;
Na.sub.2HPO.sub.4/citric acid pH 6.5) to OD.sub.650=0.5. Cells were
harvested by centrifugation and resuspended in 100 ml defined
medium. To 2 ml of washed cells was added 10 microlitres, 20
microlitres, 40 microlitres, 80 microlitres or 160 microlitres of
the mutagen stock solution. The cells were then incubated at
30.degree. C., 200 rpm for 30 rain. One ml of mutated cells was
washed twice with 1 ml sterile distilled water and finally
resuspended in 1 ml YEP. The percentage of cells that survived the
mutagenic treatment was assessed by spreading an aliquot of each
mutagenic reaction onto YEP, 2% (w/v) sucrose plates. Mutagen stock
solutions were prepared as follows.
N-methyl-N-nitro-N-nitrosoguanidine (NTG) was dissolved in ethanol
at 5 mg/mL; 4 nitroquinoline N-oxide (NQO) was resuspended in
acetone at 10 mg/mL and then diluted 1 in 100 to 0.1 mg/mL with
K.sub.2HPO.sub.4/KH.sub.2PO.sub.4 (pH 7.0); 1,2,7,8-diepoxyoctane
(DEO) and ethyl methanesulphonate (EMS) were both supplied as
liquids (Sigma) and were used without dilution.
[0594] After mutagenesis, a S. cerevisiae strain was identified
with a higher level of production of a recombinant protein,
compared to its ancestral strain (data not shown).
EXAMPLE 2
[0595] The expression of genes in the strain identified in Example
1 was compared to the expression of genes in the ancestral strain
from which it was derived (i.e. the ancestral strain displays lower
levels of production of a recombinant protein).
[0596] The comparison was made by using microarray analysis. Yeast
cells to be analysed were grown in 100 ml defined medium (0.65%
(w/v) YNB; 2% (w/v) dextrose; Na.sub.2HPO.sub.4/citric acid pH 6.5)
to OD.sub.600=2.0. The cells were immediately harvested by
centrifugation and frozen by immersion in liquid nitrogen. RNA
suitable for microarray analysis was prepared by disruption of the
cells using a micro dismembrator (Braun Melsungen, Germany) all as
described by Jones et al, 2003, Physiol. Genomics, 16, 107-118.
cDNA synthesis, labelling, hybridisation to high-density
oligonucleotide arrays (Affymetrix-Yeast S98) and scanning were
carried out as described by protocols provided by the manufacturer
(Affymetrix Inc, USA). The subsequent data was analysed using the
MAS 5.1 and DTM 3.0 software programs (Affymetrix Inc, USA).
[0597] Genes identified as being up-regulated in the strain
identified in Example 1, compared to the ancestral strain,
include--
TABLE-US-00102 TABLE 1 Gene Fold change Gene Fold change JEM1 2.63
EUG1 3.68 LHS1 2.40 MPD1 2.37 SCJ1 1.81 MPD2 1.51 KAR2 1.24 EPS1
1.10 SIL1 4.5 PDI1 1.22 FKB2 1.62 DER1 2.64 SSA3 2.61 DER3 1.67
SSA4 1.83 HRD3 1.82 SSE2 2.31 UBC7 1.33 ECM10 5.65 DOA4 1.91 ERO1
2.66 HAC1 2.05 ERV2 1.73
[0598] It will be recognised that none of SSA1, SSA2, SSE1, SSB1,
SSB2, MDJ1 or MDJ2 were identified as being over-expressed in the
strain identified in Example 1. However, these helper proteins have
been included in the present invention as a result of their
functional association to the helper proteins whose genes have been
identified as being upregulated in the strain isolated in Example
1. For example, the genes encoding SSA3, SSA4 and SSB2 have all
been identified as being over-expressed; SSA1, SSA2, SSE1, SSB1 and
SSB2 are functional equivalents of these helper proteins and so it
is anticipated that over-expression of the genes encoding any of
SSA1, SSA2, SSE1, SSB1 or SSB2 would cause the same phenotype as
the over-expression of the genes encoding any of SSA3, SSA4 or
SSB2. Similarly the gene encoding ECM10 has been identified as
being over-expressed; MDJ1 and MDJ2 are functional equivalents of
ECM10 and so it is anticipated that the over-expression of either
of the genes encoding MDJ1 or MDJ2 would cause the same phenotype
as the over-expression of the gene encoding ECM10.
EXAMPLE 3
[0599] The example describes the vector construction and yeast
transformation for the overexpression of the representative helper
proteins LHS1, SLS1, JEM1 and SCJ1.
TABLE-US-00103 TABLE 2 Primers used Primer name Sequence (5'-3') HO
5' ForNotIBbsI GCATGCGGCCGCCCGAAGACCCTACACAGGGCTTAAGGGC HO 5'
RevBsiWIMluI CCACGCGTCGTACGGGATTGCTGCTTATGAGGATA HO 3' ForMluIEcoRI
ACGCGTGAATTCAAAAAGGGAACCCGTATATTTCAGC HO 3' RevBbsIClaI
TATCGATAGTCTTCCTAATATACACATTTTAGCAGATGC pBST HO Poly For
GCATGCATACGCGTCACGCATGTGCCTCAGCGGCCGGCCGGCGCCGGGCCCC
GGACCGCCTGCAGGCTCGAGTTAATTAAGTTTAAACGAATTCGCATGCAT pBST HO Poly Rev
ATGCATGCGAATTCGTTTAAACTTAATTAACTCGAGCCTGCAGGCGGTCCGG
GGCCCGGCGCCGGCCGGCCGCTGAGGCACATGCGTGACGCGTATGCATGC Ycplac33 Poly
For CTAGATTGGATCCCTAGTCTAGGTTTAAACTAGCGATTCACCTAGGTGCTAG GAATTCTAGC
Ycplac33 Poly Rev
GCTAGAATTCCTAGCACCTAGGTGAATCGCTAGTTTAAACCTAGACTAGGGA TCCAATCTAG
LHS1forOverlap CACAATATTTCAAGCTATACCAAGCATACAATCAACTATCTCATATA
CAATGCGAAACGTTTTAAGGCT LHS1revBbvCI
GCATGCTGAGGGTGCCACTATAATATTAATGTGC SLS1forOverlap
CACCAACACACACAAAAAACAGTACTTCACTAAATTTACACACAAA
ACAAAATGGTCCGGATTCTTCCCAT SLS1revNarI
GCATGGCGCCCCACGGCAGGGCAGTTGGCAC JEM1forOverlap
CAGATCATCAAGGAAGTAATTATCTACTTTTTACAACAAATATAAAA
CAATGATACTGATCTCGGGATAC JEM1revRsrII
CGATCGGTCCGAGGGAAATAAGGCAGATCAAAG SCJ1forOverlap
CACGCTTACTGCTTTTTTCTTCCCAAGATCGAAAATTTACTGAATTAA
CAATGATTCCAAAATTATATATAC SCJ1revXhoI GCATCTCGAGGACTTTGAGACCTGTGATC
ADH1promForAleI CGATCACCGATGTGGTTGTTTCCGGGTGTACAATATGG
ADH1promRevOverlap CCTATAGCAACAAAAGCTGTTAAAAATAAAAGCCTTAAAACGTTTCG
CATTGTATATGAGATAGTTGATTG PGK1promForPspOMI
GCATGGGCCCAGATTCCTGACTTCAACTCAAG PGK1promRevOverlap
GGCAAAATAACGCTATACACTAAAAGACAGTATCCCGAGATCAGTAT
CATTGTTTTATATTTGTTGTAAAAAC TDH1promForFseI
GCATGGCCGGCCACCATATGGAGGATAAGTTGG TDH1promRevOverlap
CTAATTTCGAAGATAGGGCGCTCAAAATTATGGGAAGAATCCGGACC
ATTTTGTTTTGTGTGTTTTAAATC TEF1promForSbfI
CGGTAGTACCTGCAGGAAGCAACAGGCGCGTTGGAC TEF1promRevOverlap
GGCAACAACAATAAAGATAGTATCAAATGTATATATAATTTTGGAAT
CATTTTGTAATTAAAACTTAGATTAGATTGC URA3forPac1
CTAGAGTTAATTAAGTTTCAATTCAATTCATC URA3revPme1
GCCTGAGTTTAAACGTTTTCTTTCCAATTTTT
[0600] pBST HO regions: HO regions were amplified by PCR from
BY4741 (Brachmann et al., 1998, Yeast, 30; 14(2):115-32) genomic
DNA using the primers shown in Table 2. Fast Start High Fidelity
PCR system (Roche) was used with the conditions as recommended: 50
.mu.L final volume containing 0.2 mM dNTPs, 1.8 mM MgCl.sub.2, 0.4
.mu.M forward and reverse primers, 100 ng template genomic DNA, 2.5
U polymerase and H.sub.2O to volume. Cycling conditions: 95.degree.
C. for 2 mins followed by 35 cycles of 95.degree. C. 30 s,
60.degree. C. 30 s, 72.degree. C. 1 min and 72.degree. C. 7 reins
for final elongation.
[0601] Fragments were gel extracted from a 1% (w/v) agarose TAE gel
using the GeneClean III kit (Q-bio Gene). Purified DNA was digested
with the appropriate enzymes, NotI and MluI for HO 5' region, MluI
and ClaI for HO 3' region. pBST+ (WO99/00504) was digested with
NotI and ClaI. Fragments were purified as above. A three way
ligation was performed using a Rapid Ligation Kit (Roche) as per
manufacturers instructions. Ligations were transformed into the E.
coli strain DH5.alpha.. Diagnostic restriction digests were
performed on mini-prep DNA to confirm the ligation was successful.
The plasmid map is shown in FIG. 1.
[0602] Polylinkers: To facilitate the cloning of the helper genes a
polynucleotide linkers were incorporated into pBST+HO regions (FIG.
1) and into YCplac33 (Gietz and Sugino, 1988, Gene, 74,
527-534).
[0603] Complementary single stranded oligonucleotides were annealed
as follows: 1 .mu.L of a 100 .mu.M solution of each oligo (Poly For
and Poly Rev, Table 2) was added into a 50 .mu.L total volume
containing 10.times. restriction buffer (Roche Buffer H for pBST HO
polylinker, Buffer B for YCplac33 polylinker). Samples were placed
into a PCR machine and heated to 98.degree. C. for 4 mins. Samples
were then held for 1 min with the temperature dropping 1.degree. C.
every cycle down to 30.degree. C. The annealed polylinkers were
then digested by addition of the appropriate restriction enzyme
(MluI, EcoRI for pBST HO polylinker, BamHI, EcoRI for YCplac33
polylinker). Digested polylinkers were gel extracted as previously
and ligated into the corresponding vector digests. Incorporation of
polylinkers was confirmed by linearising plasmids with all
restriction sites present in polylinkers. Vectors produced are
shown as FIGS. 2 and 3 respectively.
[0604] Production of promoter/open reading frame constructs: All
four open reading frames (ORFs) and promoters were amplified by
PCR, from the genomic DNA of an AH22 derivative, using Vent
polymerase (NEB). Reactions were setup as per manufacturers
instructions with an annealing temperature of 50.degree. C. All
fragments were gel extracted and resuspended in 5 .mu.L of water. 1
.mu.L was run on gel to check fragment presence and quantity.
[0605] Promoters and ORFs were joined according to the method of
Shevchuk et al. (Nucleic Acids Res., 2004, 32(2), e19.). 100 ng of
ORF and an equimolar amount of promoter was used in the first PCR
stage. 10 .mu.L from this was used in the second PCR stage. Primers
were added to a final concentration of 0.4 .mu.M.
[0606] Second stage PCRs were run on a 1% (w/v) agarose TAE gel and
bands extracted of the expected size (promoter+ORF length).
Extracted fragments were A-tailed using Fast Start High Fidelity
polymerase (Roche) and cloned into the Topo pCR2.1 vector
(Invitrogen). Plasmid DNA was restriction digested to confirm the
correct insert and subsequently sequenced.
[0607] Assembly of overexpression constructs: Restriction digests
were performed to release promoter/ORF constructs from Topo pCR2.1
vectors. Fragments were gel extracted and ligated into the pBST HO
polylinker vector, digested accordingly. In the first instance,
constructs were produced containing each individual promoter/ORF
and containing all four. This required subsequent rounds of plasmid
transformation, digestion and ligation. The vector containing all
four promoter/ORFs is shown in FIG. 4.
[0608] For insertion of promoter/ORF constructs into the
centromeric vector, YCplac33 polylinker, a PmeI/AleI digest was
performed on pBST HO POLY (FIG. 4) containing the required
promoter/ORFs, and YCplac33 polylinker. The fragment released from
pBST HO POLY was ligated with the digested YCplac33 polylinker
vector. The vector containing all four promoter/ORFs is shown in
FIG. 5.
[0609] Insertion of URA3 marker into pBST HO POLY: The URA3 marker
was amplified by PCR from the vector YCp50 (Rose et al., 1987,
Gene, 60, 237-243) using Fast Start High Fidelity polymerase
(Roche) with an annealing temperature of 50.degree. C. The fragment
was gel extracted, digested with PacI/PmeI and ligated into each
pBST HO POLY vector containing the required promoter/ORFs (also
PacI/PmeI digested). It is important the URA3 fragment be
introduced last as it contains sites for restriction enzymes used
elsewhere in construction of the plasmid. The vector produced
containing all four promoter/ORFs is shown in FIG. 6.
[0610] Chromosomal integration: The helper gene constructs were
integrated into the genome of a S. cerevisiae host cell as follows.
The vector pBST HO POLY URA3 COMP (FIG. 6) was digested with NotI
and SacII. Approximately 2-3 .mu.g of the required fragment was gel
extracted and used to transform a ura3 derivative of AH22 [pAYE329]
using a yeast transformation kit (Sigma). Transformations were
plated onto minimal media and incubated at 30.degree. C. until
colonies appeared. The construction of plasmid pAYE329 is described
in Sleep et al., 1990, Gene, 101, 89-96. A ura3 auxotrophic mutant
of the AH22 derivative was created by 5-fluoro-orotic acid
selection as described by Boeke et al, 1987, Methods Enzymol., 154,
164-175.
[0611] Alternatively, the helper gene constructs may be introduced
on a centromeric vector. For the YCplac33 based-vectors, 500 ng of
plasmid DNA may be used to transform a S. cerevisiae host cell as
above.
EXAMPLE 4
[0612] This example describes a modified protocol for vector
construction and yeast transformation for the overexpression of the
representative helper proteins LHS1, SIL1; JEM1 and SCJ1.
TABLE-US-00104 TABLE 3 Primers used Primer Sequence (5'-3') -
Regions underlined indicate restriction enzyme name Product
cleavage sites and are followed by the name of the cleaving enzyme.
A01 H0 5' region
GCATGCGGCCGC(NotI)CCGAAGAC(BbsI)CCTACACAGGGCTTAAGGGC A02
CCACGCGT(MluI)CGTACG(BsiWI)GGATTGCTGCTTATGAGGATA A03 HO 3' region
ACGCGT(MluI)GAATTC(EcoRI)AAAAAGGGAACCCGTATATTTCAGC A04
TATCGAT(ClaI)AGTCTTC(BbsI)CTAATATACACATTTTAGCAGATGC A05 pTPA02
GCATGCATACGCGT(MluI)CACGCATGTGCCTCAGC(BbvCI)GGCCGGCC poly-linker
(FseI)GGCGCC(NarI)GGGCCC(PspOMI)CGGACCG(RsrII)CCTGCAGG(SbfI)
CTCGAG(XhoI)TTAATTAA(PacI)GTTTAAAC(PmeI)GAATTC(EcoRI)GCA TGCAT A06
ATGCATGCGAATTC(EcoRI)GTTTAAAC(PmeI)TTAATTAA(PacI)CTCGA
G(XhoI)CCTGCAGG(SbfI)CGGTCCG(RsrII)GGGCCC(PspOMI)GGCGCC(NarI)
GGCCGGCC(FseI)GCTGAGG(BbvCI)CACATGCGTGACGCGT(MluI)AT GCATGC A07
ACT1 CTAGGTAACTTAATTAA(PacI)GGGTAAGCTGCCACAGCA A08 promoter
CTACGTACTCTAGA(XbaI)TGTTAATTCAGTAAATTTTC A09 ACT1
CTAGACTCTAGA(XbaI)TCTCTGCTTTTGTGCGCG A10 terminator
CATGCTACGTTTAAAC(PmeI)GATGATCATATGATACAC A11 URA3 region
CTAGAGTTAATTAA(PacI)GTTTCAATTCAATTCATC A12
GCCTGAGTTTAAAC(PmeI)GTTTTCTTTCCAATTTTT A13 pTPA05
CTAGATTGGATCCCTAGTCTAGGTTTAAACTAGCGATTCACCTAGGTG poly-linker
(AleI)CTAGGAATTCTAGC A14
GCTAGAATTCCTAGCACCTAGGTG(AleI)AATCGCTAGTTTAAACCTAG
ACTAGGGATCCAATCTAG C01 LHS1 ORF/
CACAATATTTCAAGCTATACCAAGCATACAATCAACTATCTCATATAC terminator
AATGCGAAACGTTTTAAGGCT C02 GCATGCTGAGG(BbvCI)GTGCCACTATAATATTAATGTGC
C03 SIL1 ORF/ CTAGATCTCTAGA(XbaI)ATGGTCCGGATTCTTCC C04 terminator
GCATGGCGCC(NarI)CCACGGCAGGGCAGTTGGCAC C05 JEM1 ORF/
CTAGATCTCTAGA(XbaI)ATGATACTGATCTCGGG C06 terminator
CGATCGGTCCG(RsrII)AGGGAAATAAGGCAGATCAAAG C07 SCJ1 ORF/
CACGCTTACTGCTTTTTTCTTCCCAAGATCGAAAATTTACTGAATTAA terminator
CAATGATTCCAAAATTATATATAC C08 GCATCTCGAG(XhoI)GACTTTGAGACCTGTGATC
C09 ADH1 CGATCACCGATGTG(AleI)GTTGTTTCCGGGTGTACAATATGG C10 promoter
CCTATAGCAACAAAAGCTGTTAAAAATAAAAGCCTTAAAACGTTTCG
CATTGTATATGAGATAGTTGATTG C11 PGK1
GCATGGGCCC(PspOMI)AGATTCCTGACTTCAACTCAAG C12 promoter
GATCTAGTCTAGA(XbaI)TGTTTTATATTTGTTGTAA C13 TDH1
GCATGGCCGGCC(FseI)ACCATATGGAGGATAAGTTGG C14 promoter
ACCTAGTCTAGA(XbaI)TTTGTTTTGTGTGTAAATTTAG C15 TEF1
CGGTAGTACCTGCAGG(SbfI)AAGCAACAGGCGCGTTGGAC C16 promoter
GGCAACAACAATAAAGATAGTATCAAATGTATATATAATTTTGGAAT
CATTTTGTAATTAAAACTTAGATTAGATTGC C17 HAC1 ORF
CTAGTCTCTAGA(XbaI)ATGGAAATGACTGATTTTGAAC C18
CTAGTCTAGA(XbaI)TCATGAAGTGATGAAGAAATC
[0613] Construction of pTPA01: 5' and 3' regions of the HO open
reading frame were amplified by PCR from BY4741 (Brachmann et al.,
1998, Yeast, 30; 14(2):115-32) genomic DNA using the primers A01-02
(5') and A03-04 (3'). Fast Start High Fidelity PCR system (Roche)
was used with the conditions as recommended, as defined in Example
3, above.
[0614] Fragments were gel extracted from a 1% (w/v) agarose TAE gel
using the GeneClean III kit (Q-bio Gene). Purified DNA was digested
with the appropriate enzymes, NotI and MluI for HO 5' region, MluI
and ClaI for HO 3' region. pBST+ (WO99/00504) was digested with
NotI and ClaI. Fragments were purified as above. A three-way
ligation was performed using a Rapid Ligation Kit (Roche) as per
manufacturers instructions. Ligations were transformed into the E.
coli strain DH5.alpha.. Diagnostic restriction digests were
performed on mini-prep DNA to confirm the ligation was successful.
The plasmid map of TPA01 is shown in FIG. 7.
[0615] Polylinkers: To facilitate the cloning of the helper genes a
polynucleotide linker was incorporated into pTPA01 (FIG. 7) and
into YCplac33 (Gietz and Sugino, 1988, Gene, 74, 527-534).
[0616] Complementary single stranded oligonucleotides were annealed
as follows: 1 .mu.L of a 100 .mu.M solution of each oligo (A05-06
and A13-14) was added into a 50 .mu.L, total volume containing
10.times. restriction buffer (Roche Buffer H for pTPA01 polylinker,
Buffer B for YCplac33 polylinker). Samples were placed into a PCR
machine and heated to 98.degree. C. for 4 mins. Samples were then
held for 1 min with the temperature dropping 1.degree. C. every
cycle down to 30.degree. C. The annealed polylinkers were then
digested by addition of the appropriate restriction enzyme (MluI,
EcoRI for pTPA01 polylinker, BamHI, EcoRI for YCplac33 polylinker).
Digested polylinkers were gel extracted as previously and ligated
into the corresponding vector digests. Incorporation of polylinkers
was confirmed by linearising plasmids with all restriction sites
present in polylinkers. Vectors produced are shown as FIGS. 8 and
11 respectively.
[0617] Production of promoter/oven reading frame constructs: LHS1,
SIL1, JEM1 and SCJ1 open reading frames (ORFs) plus approximately
300 bp of terminator sequence (3' of ORF) and promoters were
amplified by PCR, from the genomic DNA of an AH22 derivative, using
Vent polymerase (NEB) (see Table 3 for primers used). Reactions
were setup as per manufacturers instructions with an annealing
temperature of 50.degree. C. All fragments were gel extracted and
resuspended in 5 .mu.L of water. 1 .mu.L was run on a gel to check
fragment presence and quantity.
[0618] Promoters and ORFs for LHS1 and SCJ1 were joined according
to the method of Shevchuk et al. (Nucleic Acids Res., 2004, 32(2),
e19.). 100 ng of the ORF fragment and an equimolar amount of
promoter fragment was used in the first PCR stage. 10 .mu.L from
this was used in the second PCR stage. Primers were added to a
final concentration of 0.4 .mu.M.
[0619] Second stage PCRs were run on a 1% (w/v) agarose TAE gel and
bands extracted of the expected size (promoter+ORF+terminator).
Extracted fragments were A-tailed using Fast Start High Fidelity
polymerase (Roche) and cloned into the TOPO pCR2.1 vector
(Invitrogen). Plasmid DNA was restriction digested to confirm the
correct insert.
[0620] Promoters and ORFs for SIL1 and JEM1 were digested with
restriction enzymes corresponding to sites incorporated into
primers used for PCR (see Table 3). Promoter and ORF fragments were
then joined by three way ligation with digested pTPA02.
[0621] The ACT1 promoter and terminator were amplified by PCR from
the genomic DNA of an AH22 derivative and gel extracted. Purified
fragments were digested with restriction enzymes corresponding to
sites incorporated into primers used for PCR and ligated in a three
way ligation with PacI/PmeI digested pTPA02 to create pTPA03 (FIG.
9).
[0622] The HAC1 ORF was amplified by PCR from cDNA derived from RNA
from an AH22 derivative treated with the reducing agent
dithiothreitol (DTT). The spliced form of HAC1 (HAC1.sup.i) was
identified as a 717 bp fragment and gel extracted. The extracted
fragment was then digested with XbaI and ligated into pTPA03
digested with the same enzyme. Diagnostic restriction digests were
used to confirm that the HAC1 ORF was present in the correct
orientation relative to the ACT1 promoter and terminator sequences.
The resultant plasmid pTPC01 is shown in FIG. 13.
[0623] All ORFs were sequenced and, with exception of LHS1, were
shown to contain the same sequence as that published for the strain
S288C. Repeat sequencing of multiple cloned PCR products for LHS1
confirmed that the AH22 derived clones contained a single base
change from the S288C sequence. The base change at position 1215
(relative to the first base of the start codon) results in a change
from A to C, which produces a Lys to Asn substitution at position
405.
[0624] Assembly of overexpression constructs: Restriction digests
(see Table 3) were performed to release promoter/ORF constructs
from TOPO pCR2.1 vectors. Fragments were gel extracted and ligated
into the pTPA02 vector, digested accordingly. In the first
instance, constructs were produced containing each individual
promoter/ORF and then containing all four. This required subsequent
rounds of plasmid transformation, digestion and ligation. The
vector containing all four promoter/ORFs is shown in FIG. 12.
[0625] For insertion of the various promoter/ORF constructs (with
the exception of HAC1) into the centromeric vector, pTPA05 (FIG.
11), an AleI/XhoI digest was performed on the various pTPA02 based
vectors containing the required promoter/ORFs (e.g. pTPC08 (FIG.
12) for LHS1, SIL1, JEM1 and SCJ1), and an AleI/SalI digest on
pTPA05 (FIG. 11). The various promoter/ORF fragments released were
ligated into AleI/Sa/I digested pTPA05 to create a series of
vectors including pTPC18 (FIG. 14) containing all four
promoter/ORFs.
[0626] Plasmid pTPC17 (Example 4, FIG. 15) contained the LHS1, SIL1
and JEM1 ORFs expressed from YCplac33. pTPC17 was constructed by
cloning an approximately 9.0-kb AleI-XhoI DNA fragment from pTPC07
(FIG. 16) that contained the expression cassette for the LHS1, SIL1
and JEM1 ORFs, into pTPA05 (FIG. 11) which had been digested with
AleI and SalI. The expression cassette for the LHS1, SIL1 and JEM1
ORFs was assembled in pTPA05 in a similar method to that described
for pTPC08 (FIG. 12), but using the promoter/ORF constructs from
TOPO pCR2.1 vectors for LHS1, SIL1 and JEM1 expression.
[0627] For insertion of the HAC1 promoter/ORF (FIG. 13) into the
centromeric vector pTPA05, an AleI/BclI digest was performed on
pTPC01 (FIG. 13) and an AleI/BamHI digest was performed on pTPA05
(FIG. 11). The HAC1 AleI/BclI fragment released from pTPC01 was
ligated into the AleI/BamHI digested pTPA05.
[0628] The various promoter/ORF constructs comprising the YCplac33
based plasmids pTPC11, pTPC12, pTPC13, pTPC14, pTPC15, pTPC17 and
pTPC18 are shown in Table 4.
TABLE-US-00105 TABLE 4 Plasmid compositions Name Helper genes
overexpressed Promoter used YCplac33 -- -- pTPC11 HAC1.sup.i ACT1
pTPC12 SIL1 TDH1 pTPC13 LHS1 ADH1 pTPC14 JEM1 PGK1 pTPC15 SCJ1 TEF1
pTPC17 LHS1, JEM1, SIL1 As shown individually above pTPC18 LHS1,
JEM1, SIL1, SCJ1 As shown individually above
[0629] Insertion of URA3 marker into pTPA02: The URA3 marker was
amplified by PCR from the vector YCp50 as described above in
Example 3. The fragment was gel extracted, digested with PacI/PmeI
and ligated into each pTPA02 based vector containing the required
promoter/ORFs (also PacI/PmeI digested). It is important the URA3
fragment be introduced last as it contains sites for restriction
enzymes used elsewhere in construction of the plasmid.
[0630] Chromosomal integration: The helper gene constructs were
integrated into the genome of a S. cerevisiae host cell by
digestion of the vector pTPC08 (FIG. 12) with Nod and SacII and
transformation of a ura3 derivative of AH22 [pAYE329] as described
in Example 3, above.
[0631] Alternatively, the helper gene constructs may be introduced
on a centromeric vector. For the YCplac33 based-vectors, 500 ng of
plasmid DNA may be used to transform a S. cerevisiae host cell as
above.
EXAMPLE 5
[0632] Plasmids constructs were produced for the overexpression of
the genes LHS1, JEM1, SCJ1 and SIL1 as described in Example 4,
above.
[0633] The spliced form of the transcription factor HAC1 (referred
to as HAC1.sup.i) was also overexpressed using the vector series
produced. Due to the regulatory role of HAC1 within the unfolded
protein response, HAC1s was overexpressed alone, not in conjunction
with the other chaperone genes described here.
[0634] All genes were overexpressed from YCplac33 based vectors
(Table 4) and transformed into the ura3 auxotrophic mutant of the
ancestral S. cerevisiae strain (a histidine revertant of AH22)
[pAYE329] defined in Example 4, above.
[0635] Overexpression was confirmed using real time PCR. Taqman
hybridisation probes were designed to bind specifically to each
gene under investigation plus ACT1, used here as an endogenous
control. An additional probe was designed for the gene HAC1 to bind
across the exon-exon junction--resulting in binding only to the
spliced form. The proportion of HAC1.sup.i relative to total HAC1
can thus be determined.
TABLE-US-00106 TABLE 5 Taqman probe/primer sequences and binding
co-ordinates Gene Name Feature Sequence/Coordinates ACT1 Forward
primer (5'-3') CCCAGAAGCTTTGTTCCATCCTT Reverse primer (5'-3')
ATGATGGAGTTGTAAGTAGTTTGGTCAA Probe (5'-3') CAGATTCCAAACCCAAAACA
Coordinates* 795-814 LHS1 Forward primer (5'-3')
ACACTACTCAGCCCGTTACAATAGA Reverse primer (5'-3')
GTAAACTTTGCACCACCTAGATGTG Probe (5'-3') ATTTGAAGGATATGGGTATAATC
Coordinates* 789-811 SIL1 Forward primer (5'-3')
GACATGTACGAAAATGACGATACAAATCT Reverse primer (5'-3')
TCGTTTGCCCACTCTTGCA Probe (5'-3') TTTGACGACCAATTCTC Coordinates*
940-956 SCJ1 Forward primer (5'-3') GGCGCAGGTGGATTCCA Reverse
primer (5'-3') CGCCAGGACCTCCATGAC Probe (5'-3')
CATATTCGAACGGATGTTTC Coordinates* 342-361 JEM1 Forward primer
(5'-3') CCTCTCCACGCACATCGA Reverse primer (5'-3')
TGCTTGTCGAGGATTGTTTCGTAAT Probe (5'-3') TCGTTAGCTGCTGCTATCA
Coordinates* 592-610 HAC1 Forward primer (5'-3')
GAAGACGCGTTGACTTGCA Reverse primer (5'-3')
GAAATCCCTGTACTCGTCAAGAGAA Probe (5'-3') CCACGACGCTTTTGTTGC
Coordinates* 288-305 HAC1.sup.i Forward primer (5'-3')
ACAATTCAATTGATCTTGACAATTGG Reverse primer (5'-3')
TCAATTCAAATGAATCAAACCTGAC Probe (5'-3') CGTAATCCAGAAGCGCA
Coordinates* 652-668 *means probe binding coordinates, relative to
start codon
[0636] The relative standard curve method of transcript
quantification was used as described by Applied Biosystems in the
`ABI PRISM 770 Sequence Detection System: User Bulletin #2`
document. This can be downloaded from the Applied Biosystems'
website (www.appliedbiosystems.com). Equivalent technical
disclosure of a suitable quantitative RT-PCR method can be found in
Bustin, 2000, Journal of Molecular Endocrinology, 25, 169-193. This
method allows quantification of the gene of interest relative to an
endogenous control gene that is known to exhibit constant
expression across experimental conditions.
[0637] All real time PCR was carried out on cDNA derived from RNA
extracted from log phase (OD.sub.600=2) BMMD yeast cultures.
Overexpression was assessed by comparison of strains with a control
yeast strain transformed with the base vector YCplac33 and are
expressed as fold changes.
TABLE-US-00107 TABLE 6 Summary of overexpression levels achieved
Overexpression in single gene construct Overexpression in multiple
gene (Fold change vs. construct pTPC18 Gene YCplac33 control) (Fold
change vs. YCplac33 control) HAC1.sup.i 3.51 -- LHS1 22.63 23.52
JEM1 10.16 11.48 SIL1 2.03 2.36 SCJ1 15.81 16.71
[0638] As shown below in Table 6, overexpression levels vary
between the different constructs. Levels achieved range from 2.03
fold for SIL1 to 22.63 fold for LHS1.
[0639] The effect of overexpression of HAC1.sup.i, LHS1, JEM1, SIL1
and SCJ1 on the induction of the stress-related unfolded protein
response (UPR) in a host cell was investigated by measuring the
levels of HAC1.sup.i and total HAC1 transcript levels in AH22
(ura3) [pAYE329] host cells transformed with Ycplac33 (as a
negative control), pTPC11, pTPC12, pTPC13, pTPC14, pTPC15 or
pTPC18. Total HAC1 transcript levels are the sum of HAC1.sup.i
transcript levels and unspliced HAC1 transcript levels. A reduced
proportion of the level of HAC1.sup.i transcript levels compared to
total HAC1 transcript levels is indicative of reduced stress and
reduced UPR signalling.
[0640] FIG. 10 shows that individual over-expression of LHS1
(pTPC13) or JEW (pTPC14) or simultaneous over-expression of all of
LHS1, JEM1, SIL1 and SCJ1 (pTPC18) resulted a reduced proportion of
the level of HAC1.sup.i transcript levels (compared to total HAC1
transcript levels) compared to the control. This indicates that
over-expression of the above-identified helper proteins can help to
reduce stress in cultured cells and avoid the unnecessary induction
of the UPR.
EXAMPLE 6
[0641] The levels of recombinant protein production achieved by the
transformed strains described in Examples 4 and 5 (see Table 4),
above, were analysed. In this case, the recombinant protein was
recombinant human albumin ("rHA") expressed from the plasmid
pAYE329, described in Sleep et al.; 1990, Gene, 101, 89-96.
[0642] All analysis was performed on cultures grown for 5 days at
30.degree. C., 200 rpm.
[0643] Culture supernatants were run immediately on gels to prevent
any rHA proteolysis/degradation that could otherwise occur during
freezing and overnight storage at -20.degree. C. Each of the three
bands (main rHA band plus two degradation products) were quantified
by densitometry. This gives an indication of rHA production levels
and the level of proteolysis occurring in each strain. The
mutagenised strain identified in Example 1 was also included as a
positive control.
[0644] Results of the analysis are shown in FIG. 17. It is apparent
from a comparison of the results for the ancestral strain
expressing recombinant albumin from pAYE329/YCplac33 ("YCplac33")
and the mutagenised strain identified in. Example 1 as possessing
increased recombinant protein production ("+ve control") that the
mutagenised strain is not only capable of producing increased
levels of rHA, but additionally displays reduced levels of rHA
degradation compared to the ancestral strain. Moreover, FIG. 17 is
particularly clear in demonstrating that strain transformed with
pTPC17 (i.e. the ancestral strain transformed to over-express LHS1,
JEM1 and SIL1) also displays reduced levels of rHA degradation
compared to the untransformed ancestral strain.
[0645] Further characterisation of the effect of the defined
transformations is possible in view of the analysis of the SDS-PAGE
gel by densitometry, the results of which are present in Table 7,
below, and FIGS. 18 and 19.
TABLE-US-00108 TABLE 7 Comparison of rHA levels, as percentage of
YCplac33 control production levels. In the third column, the rHA
production levels have been normalised (based on culture optical
density readings) to account for different growth rates observed
between transformants. rHA production, Overexpression as % of
YCplac33 control plasmid Not normalised by OD Normalised by OD
pTPC11 164.26 139.2 pTPC12 102.51 101.7 pTPC13 122.42 115.1 pTPC14
177.85 170.4 pTPC15 86.37 103.4 pTPC17 132.85 116.3 pTPC18 102.65
96.0 +ve control 383.44 369.0
[0646] Table 7, above, and FIGS. 18 and 19, show that the
individual overexpression of HAC1, LHS7, JEM1, SIL1 and SCJ1
results in an increase in rHA production, on a per cell basis (i.e.
when results are normalised by culture OD). However, the negative
growth effect of SCJ1 overexpression resulted in an overall
reduction of rHA production on a per culture basis (i.e. when
results are not normalised by culture OD).
[0647] The overexpression of JEM1 alone had the largest measured
effect on rHA production.
[0648] However, as will be apparent from FIG. 17, the strains that
individually expressed HAC1, LHS1, JEM1, SIL1 and SCJ1 still
demonstrated relatively high levels of rHA degradation, comparable
to the ancestral strain and higher than the to mutagenised strain
identified in Example 1. By contrast, cells that simultaneously
over-express LHS1, JEM1 and SIL1 demonstrate increased rHA
productivity and a concomitant reduction in rHA degradation,
comparable with the mutagenised strain identified in Example 1.
This is further demonstrated in FIG. 20. In fact, FIG. 20 shows
that several of the strains tested show lower levels of degradation
compared to the ancestral strain, but this reduction is
particularly pronounced in strain transformed with pTPC17.
EXAMPLE 7
[0649] This example describes the increased secretion of a
recombinant transferrin mutant by over-expression of LHS1, JEM1 and
SIL1 from the centromeric vector pTPC17 in a Saccharomyces
cerevisiae strain containing a 2-micron plasmid encoding the PDI1
gene.
[0650] A S. cerevisiae strain, the "control strain" as used in WO
2005/061718 and WO 2005/061719 was used to generate a ura3 mutant
derivative, referred to herein as "control strain (ura3)" by random
mutagenesis and selection on 5-fluoro-orotic acid plates (Boeke et
al., 1984, op. cit.).
[0651] The S. cerevisiae control strain was transformed to leucine
prototrophy with pDB3213 (FIG. 21) and the control strain (ura3)
was co-transformed to both leucine and uracil prototrophy with
plasmids pTPC17 (FIG. 15) and pDB3213. Transformation was by a
modified lithium acetate method (Sigma yeast transformation kit,
YEAST-1, protocol 2 (Elble, R, 1992, Biotechniques, 13, 18-20; Ito
et al., 1983, op. cit.). Transformants were selected on BMMD-agar
plates, and subsequently patched out on BMMD-agar plates.
[0652] The construction of pTPC17 is described in Example 4.
[0653] Plasmid pDB3213 is similar to pDB2929 (WO 2005/061718,
Example 1 and FIG. 12), and contains a NotI expression cassette for
a non-glycosylated transferrin cloned into pDB2690 (WO 2005/061718,
Example 1 and FIG. 6). The NotI expression cassette of pDB3213
contains an alternative codon for Leucine-505 in mature transferrin
that is the CTG codon (11% codon usage in S. cerevisiae) compared
to the CTG codon (6% codon usage in S. cerevisiae) present in
pDB2929, a KEX2-independent leader sequence (derived from the
HSA-pre leader sequence) and mutations within the N-linked
glycosylation sites (-N-X-S/T-) that prevent glycosylation of
residues N413 and N611.
[0654] Transformants of each strain were inoculated into 10 mL BMMD
and 10 mL YEPD in 50 mL shake flasks and incubated in an orbital
shaker at 30.degree. C., 200 rpm for 4-days. Culture supernatants
were harvested and the recombinant transferrin titres compared by
rocket immunoelectrophoresis (FIG. 22). The results indicated that
the recombinant transferrin titres in supernatants of both the YEPD
and BMMD shake flask cultures were higher when pTPC17 was present.
Furthermore, in high cell density fed batch fermentation the
recombinant transferrin titres from control strain (ura3) [pTPC17
pDB3213] was 1.7 g/L compared to only 0.9 g/L for control strain
[pDB3213]. Therefore, over-expression of LHS1, JEM1 and SIL1 from
the centromeric plasmid pTPC17 had approximately doubled the
quantity of the recombinant transferrin product secreted from the
S. cerevisiae strain during fermentation.
[0655] It is to be noted that pDB3213 encodes an additional copy of
PDI1, and these results suggest that over-expression of PDI1 (and
variants thereof) in conjunction with one, two or all three of
LHS1, JEM1, and SIL1 (e.g. LHS1 alone; JEM1 alone; SIL1 alone; LHS1
and JEM1; LHS1, and SIL1; JEM1, and SIL1; or LHS1, JEM1, and SIL1)
provide unexpected benefits to the production of a desired protein
product.
EXAMPLE 8
[0656] This example shows increased secretion of recombinant
albumin ("rHA") by over-expression of LHS1, JEM1 and SIL1 from the
centromeric vector pTPC17 in a Saccharomyces cerevisiae strain.
[0657] Construction of plasmid pDB2243 containing the NotI rHA
expression cassette, incorporating the HSA/MF.alpha.-1 fusion
leader sequence, as taught in WO 90/01063, is described in WO
00/44772 (see WO 00/44772, FIG. 6). The rHA expression
disintegration vector pDB2244 (FIG. 23) was created by ligating the
NotI expression cassette from pDB2243 into NotI cut pSAC35 (Sleep
et al, 1991, Bio/Technology 9, 183-187 and EP 431 880) to generate
the plasmid pDB2244 in which the direction of rHA transcription is
in the same orientation as that of the LEU2 gene as described in WO
00/44772.
[0658] Construction of plasmid pDB2283 containing a NotI rHA
expression cassette, incorporating the invertase leader sequence,
was accomplished by replacing the 1.21-kb BfrI-XbaI fragment in
pDB2243, comprising the HSA/MF.alpha.-1 fusion leader sequence and
part of the human albumin cDNA, with a 1.07-kb blunt end-XbaI
fragment from mp 19.7 (EP-A-248 637) and a synthetic double
stranded oligonucleotide linker of the following structure--
TABLE-US-00109 1 gagtccaatt agcttcatcg ccaataaaaa aacaagctaa
acctaattct ctcaggttaa tcgaagtagc ggttattttt ttgttcgatt tggattaaga
HindIII -+---- 51 aacaagcaaa gatgaagtgg gtaagcttaa cctaattcta
acaagcaaag ttgttcgttt ctacttcacc cattcgaatt ggattaagat tgttcgtttc
101 atgcttttgc aagccttcct tttccttttg gctggttttg cagccaaaat
tacgaaaacg ttcggaagga aaaggaaaac cgaccaaaac gtcggtttta
>>......................Invertase......................> m
l l q a f l f l l a g f a a k 151 atctgca tagacgt >....>>
Invertase i s a
which was formed by annealing two complementary single stranded
oligonucleotides with the sequences
TABLE-US-00110 5'TTAAGAGTCCAATTAGCTTCATCGCCAATAAAAAAACAAGCTAAACCT
AATTCTAACAAGCAAAGATGAAGTGGGTAAGCTTAACCTAATTCTAACAA
GCAAAGATGCTTTTGCAAGCCTTCCTTTTCCTTTTGGCTGGTTTTGCAGC CAAAATATCTGCA3';
and 5'TGCAGATATTTTGGCTGCAAAACCAGCCAAAAGGAAAAGGAAGGCTTG
CAAAAGCATCTTTGCTTGTTAGAATTAGGTTAAGCTTACCCACTTCATCT
TTGCTTGTTAGAATTAGGTTTAGCTTGTTTTTTTATTGGCGATGAAGCTA ATTGGACTC3'.
[0659] Plasmid mp 19.7 (EP-A-248 637) was digested to completion
with XhoI, phenol/chloroform extracted and ethanol precipitated.
The recovered DNA was then blunt ended with the Klenow fragment of
E. coli DNA polymerase I to remove the XhoI overhang,
phenol/chloroform extracted, and ethanol precipitated. The
recovered DNA was digested to completion with XbaI. The digestion
products were resolved by agarose gel electrophoresis and the
1.07-kb blunt end-XbaI mp 19.7 fragment recovered using the
GeneClean III kit (Q-bio Gene).
[0660] The rHA expression disintegration vector pDB2286 (FIG. 24)
was created by ligating the Nod expression cassette from pDB2283
into NotI cut pSAC35 (Sleep et al, 1991, Bio/Technology 9, 183-187
and EP 431 880).
[0661] Construction of plasmid pDB2284 containing a Nod rHA
expression cassette, incorporating the MF.alpha.-1 leader sequence,
was accomplished by replacing the 1.21-kb BrfI-XbaI fragment in
pDB2243, comprising the HSA/MF.alpha.-1 fusion leader sequence and
part of the human albumin cDNA, with a 1.07-kb blunt end-XbaI
fragment from mp 19.7 (EP-A-248 637) and a synthetic double
stranded phosphorylated oligonucleotide linker of the
structure--
TABLE-US-00111 1 ttaagagtcc aattagcttc atcgccaata aaaaaacaaa
ctaaacctaa ctcagg ttaatcgaag tagcggttat ttttttgttt gatttggatt PstI
------+ 51 ttctaacaag caaagatgag atttccttca atttttactg cagttttatt
aagattgttc gtttctactc taaaggaagt taaaaatgac gtcaaaataa
>>.............MFalpha..............> m r f p s i f t a v
l 101 cgcagcatcc tccgcattag ctgctccagt caacactaca acagaagatg
gcgtcgtagg aggcgtaatc gacgaggtca gttgtgatgt tgtcttctac > MFalpha
> f a a s s a l a a p v n t t t e d 151 aaacggcaca aattccggct
gaagctgtca tcggttactc agatttagaa tttgccgtgt ttaaggccga cttcgacagt
agccaatgag tctaaatctt > MFalpha > e t a q i p a e a v i g y s
d l e 201 ggggatttcg atgttgctgt tttgccattt tccaacagca caaataacgg
cccctaaagc tacaacgaca aaacggtaaa aggttgtcgt gtttattgcc > MFalpha
> g d f d v a v l p f s n s t n n 251 gttattgttt ataaatacta
ctattgccag cattgctgct aaagaagaag caataacaaa tatttatgat gataacggtc
gtaacgacga tttcttcttc > MFalpha > g l l f i n t t i a s i a a
k e e HindIII -+---- 301 gggtaagctt ggataaaaga cccattcgaa
cctattttct >......MFalpha.....>> g v s l d k r
formed by annealing complementary six single stranded
oligonucleotides with the sequences
TABLE-US-00112 5'TTAAGAGTCCAATTAGCTTCATCGCCAATAAAAAAACAAACTAAACCT
AATTCTAACAAGCAAAGATGAGATTTCCTTCAATTTTTACTGCAGTTTTA 3';
5'TTCGCAGCATCCTCCGCATTAGCTGCTCCAGTCAACACTACAACAGAA
GATGAAACGGCACAAATTCCGGCTGAAGCTGTCATCGGTTACTCAGATTT AGAAGGGGATTT3';
5'CGATGTTGCTGTTTTGCCATTTTCCAACAGCACAAATAACGGGTTATT
GTTTATAAATACTACTATTGCCAGCATTGCTGCTAAAGAAGAAGGGGTAA
GCTTGGATAAAAGA3';
5'TCTTTTATCCAAGCTTACCCCTTCTTCTTTAGCAGCAATGCTGGCAAT
AGTAGTATTTATAAACAATAACCCGTTATTTGTGCTGTTGGAAAATGGCA AAAC3';
5'AGCAACATCGAAATCCCCTTCTAAATCTGAGTAACCGATGACAGCTTC
AGCCGGAATTTGTGCCGTTTCATCTTCTGTTGTAGTGTTGACTGGAGCAG CTAATGCGGAGG3';
and 5'ATGCTGCGAATAAAACTGCAGTAAAAATTGAAGGAAATCTCATCTTTG
CTTGTTAGAATTAGGTTTAGTTTGTTTTTTTATTGGCGATGAAGCTAATT GGACTC3'.
[0662] Plasmid mp 19.7 (EP-A-248 637) was digested to completion
with XhoI, phenol/chloroform extracted and ethanol precipitated.
The recovered DNA was then blunt ended with the Klenow fragment of
E. coli DNA polymerase I to remove the XhoI overhang,
phenol/chloroform extracted, and ethanol precipitated. The
recovered DNA was digested to completion with XbaI. The digestion
products were resolved by agarose gel electrophoresis and the
1.07-kb blunt end-XbaI mp 19.7 fragment recovered using the
GeneClean III kit (Q-bio Gene).
[0663] The rHA expression disintegration vector pDB2287 (FIG. 25)
was created by ligating the NotI expression cassette from pDB2284
into NotI cut pSAC35 (Sleep et al, 1991, Bio/Technology 9, 183-187
and EP 431 880).
[0664] The ura3 auxotrophic mutant of the AH22 histidine revertant
described in Example 4 was co-transformed to both leucine and
uracil prototrophy with plasmids pDB2244 and YCplac33, or pDB2244
and pTPC17, or pDB2286 and YCplac33, or pDB2286 and pTPC17, or
pDB2287 and YCplac33, or pDB2287 and pTPC17. Transformation was by
a modified lithium acetate method (Sigma yeast transformation kit,
YEAST-1, protocol 2 (Elble, 1992, op. cit.; Ito et al, 1983, op.
cit.). Transformants were selected on BMMD-agar plates, and
subsequently patched out on BMMD-agar plates.
[0665] Two transformants for each strain were inoculated into 10 mL
BMMD in 50 mL shake flasks and incubated in an orbital shaker at
30.degree. C., 200 rpm for 4-days. Culture supernatants were
harvested and the recombinant human albumin (rHA) titres compared
by SDS-PAGE (FIG. 26 A-C) and densitometric analysis (FIG. 26 D).
The results are summarised in Table 8, below.
TABLE-US-00113 TABLE 8 Increased rHA secretion by overexpression of
SIL1, LHS1 and JEM1 (pTPC17) from three distinct leader sequences.
Average percentage increase in rHA secretion by pTPC17 Expression
plasmid versus YCplac33 transformation pDB2244 29.1 pDB2286 16.7
pDB2287 14.5
[0666] The results indicated that the rHA titres were increased by
transformation with pTPC017 relative to the control plasmid
YCplac33. Increases in rHA titres varied between the different
expression constructs in the range of 14.5-29.1% demonstrating the
beneficial effect of LHS1, JEM1 and SIL1 on rHA secretion was not
restricted to a specific secretory leader sequence. Thus, for
example, it is clear that the beneficial effect of LHS1, JEM1 and
SIL1 on rHA secretion was not restricted by features of the leader
sequence at the amino acid or DNA sequence level, or by
configuration (pre or pre-pro) or whether or not the secretory
leader sequence contained N-linked glycosylation sites.
EXAMPLE 9
[0667] This example describes the increased secretion of
recombinant granulocyte macrophage colony stimulating factor
(GM-CSF) from a 2-micron based plasmid by over-expression of LHS1,
JEM1 and SIL1 from the centromeric vector pTPC17.
[0668] A cDNA for human GM-CSF was obtained from plasmid pBBG12
(R&D Systems Europe Ltd.) cloned between the HindIII and EcoRI
sites of the pUC18 polylinker. The DNA sequence of the human GM-CSF
cDNA (FIG. 27) incorporated an N-terminal Met codon.
[0669] Oligonucleotides SINK1 and SINK 2 were synthesised to
construct a linker which would reconstruct the HSA/MF.alpha.-1
fusion leader as taught in WO 90/01063, coupled to GM-CSF up to the
BstEII site.
TABLE-US-00114 SINK1: 5'GTACCAAGCTTTATTTCCCTTCTTTTTCTCTTTAGCTCGGC
TTATTCCAGGAGCTTGGATAAAAGAGCACCCGCCCG3' SINK2:
5'GTGACCGGGCGGGTGCTCTTTTATCCAAGCTCCTGGAATAA
GCCGAGCTAAAGAGAAAAAGAAGGGAAATAAAGCTTG3'
[0670] A 380 bp BstEII/BamHI GMCSF fragment was isolated from
pBBG12 and ligated into pUC19 Asp718/BstEII along with the
Asp718/BstEII SINK1/2 linker above, to create pDB2095. Accordingly,
the GM-CSF cDNA, linked to the HSA/MF.alpha.-1 fusion secretion
leader, was available on a HindIII fragment suitable for subcloning
into pAYE441 (as described in WO 2004/009819, Example 1 and FIG. 5)
to create pDB2102 in which the GM-CSF cDNA was now present on a
NotI expression cassette, comprising the PRB1 promoter, the
HSA/MF.alpha.-1 fusion secretion leader and the ADH1 terminator.
The GM-CSF NotI expression cassette was isolated and subcloned into
pSAC35 (Sleep et al, 1991; Biotechnology (NY), 9, 13 and EP 431
880) linearised with NotI to create plasmid pDB2109 (FIG. 28).
[0671] The S. cerevisiae Control Strain (ura3), as described above
in Example 7, was co-transformed to both leucine and uracil
prototrophy with plasmids pDB2109 (FIG. 28) and either YCplac33 or
pTPC17 (FIG. 15). Transformation was by a modified lithium acetate
method (Sigma yeast transformation kit, YEAST-1, protocol 2 (Elble,
1992, op. cit.; Ito et al., 1983, op. cit.). Transformants were
selected on BMMD-agar plates, and subsequently patched out on
BMMD-agar plates.
[0672] Transformants of each strain were inoculated into 10 mL BMMD
in 50 mL shake flasks and incubated in an orbital shaker at
30.degree. C., 200 rpm for 4-days. Culture supernatants were
harvested and the recombinant GM-CSF titres compared by SDS-PAGE
and densitometric analysis (FIGS. 29 A and B). The results of the
densitometric analysis are also provided in Table 9, below.
TABLE-US-00115 TABLE 9 Increased GM-CSF production as determined by
SDS-PAGE and densitometric analysis Control strain Control strain
(ura3) [pDB2109 YCplac33] (ura3) [pDB2109 pTPC17] Integrated
optical Integrated optical Gel lane density Gel lane density 2
45.20 6 108.36 3 72.14 7 108.41 4 71.54 8 111.73 5 74.21 9 111.30
Average 65.77 Average 109.95
[0673] The results indicated that the recombinant GM-CSF titres in
supernatants of BMMD shake flask cultures were greater than 50%
higher when pTPC17 was present.
Sequence CWU 1
1
1951645PRTSaccharomyces cerevisiae 1Met Ile Leu Ile Ser Gly Tyr Cys
Leu Leu Val Tyr Ser Val Ile Leu1 5 10 15Pro Val Leu Ile Ser Ala Ser
Lys Leu Cys Asp Leu Ala Glu Leu Gln 20 25 30Arg Leu Asn Lys Asn Leu
Lys Val Asp Thr Glu Ser Leu Pro Lys Tyr 35 40 45Gln Trp Ile Ala Gly
Gln Leu Glu Gln Asn Cys Met Thr Ala Asp Pro 50 55 60Ala Ser Glu Asn
Met Ser Asp Val Ile Gln Leu Ala Asn Gln Ile Tyr65 70 75 80Tyr Lys
Ile Gly Leu Ile Gln Leu Ser Asn Asp Gln His Leu Arg Ala 85 90 95Ile
Asn Thr Phe Glu Lys Ile Val Phe Asn Glu Thr Tyr Lys Gly Ser 100 105
110Phe Gly Lys Leu Ala Glu Lys Arg Leu Gln Glu Leu Tyr Val Asp Phe
115 120 125Gly Met Trp Asp Lys Val His Gln Lys Asp Asp Gln Tyr Ala
Lys Tyr 130 135 140Leu Ser Leu Asn Glu Thr Ile Arg Asn Lys Ile Ser
Ser Lys Asp Val145 150 155 160Ser Val Glu Glu Asp Ile Ser Glu Leu
Leu Arg Ile Thr Pro Tyr Asp 165 170 175Val Asn Val Leu Ser Thr His
Ile Asp Val Leu Phe His Lys Leu Ala 180 185 190Glu Glu Ile Asp Val
Ser Leu Ala Ala Ala Ile Ile Leu Asp Tyr Glu 195 200 205Thr Ile Leu
Asp Lys His Leu Ala Ser Leu Ser Ile Asp Thr Arg Leu 210 215 220Ser
Ile His Tyr Val Ile Ser Val Leu Gln Thr Phe Val Leu Asn Ser225 230
235 240Asp Ala Ser Phe Asn Ile Arg Lys Cys Leu Ser Ile Asp Met Asp
Tyr 245 250 255Asp Lys Cys Lys Lys Leu Ser Leu Thr Ile Ser Lys Leu
Asn Lys Val 260 265 270Asn Pro Ser Lys Arg Gln Ile Leu Asp Pro Ala
Thr Tyr Ala Phe Glu 275 280 285Asn Lys Lys Phe Arg Ser Trp Asp Arg
Ile Ile Glu Phe Tyr Leu Lys 290 295 300Asp Lys Lys Pro Phe Ile Thr
Pro Met Lys Ile Leu Asn Lys Asp Thr305 310 315 320Asn Phe Lys Asn
Asn Tyr Phe Phe Leu Glu Glu Ile Ile Lys Gln Leu 325 330 335Ile Glu
Asp Val Gln Leu Ser Arg Pro Leu Ala Lys Asn Leu Phe Glu 340 345
350Asp Pro Pro Ile Thr Asp Gly Phe Val Lys Pro Lys Ser Tyr Tyr His
355 360 365Thr Asp Tyr Leu Val Tyr Ile Asp Ser Ile Leu Cys Gln Ala
Ser Ser 370 375 380Met Ser Pro Asp Val Lys Arg Ala Lys Leu Ala Ala
Pro Phe Cys Lys385 390 395 400Lys Ser Leu Arg His Ser Leu Thr Leu
Glu Thr Trp Lys His Tyr Gln 405 410 415Asp Ala Lys Ser Glu Gln Lys
Pro Leu Pro Glu Thr Val Leu Ser Asp 420 425 430Val Trp Asn Ser Asn
Pro His Leu Leu Met Tyr Met Val Asn Ser Ile 435 440 445Leu Asn Lys
Ser Arg Ser Lys Pro His Ser Gln Phe Lys Lys Gln Leu 450 455 460Tyr
Asp Gln Ile Asn Lys Phe Phe Gln Asp Asn Gly Leu Ser Glu Ser465 470
475 480Thr Asn Pro Tyr Val Met Lys Asn Phe Arg Leu Leu Gln Lys Gln
Leu 485 490 495Gln Thr Tyr Lys Glu His Lys His Arg Asn Phe Asn Gln
Gln Tyr Phe 500 505 510Gln Gln Gln Gln Gln Gln Gln Gln His Gln Arg
His Gln Ala Pro Pro 515 520 525Ala Ala Pro Asn Tyr Asp Pro Lys Lys
Asp Tyr Tyr Lys Ile Leu Gly 530 535 540Val Ser Pro Ser Ala Ser Ser
Lys Glu Ile Arg Lys Ala Tyr Leu Asn545 550 555 560Leu Thr Lys Lys
Tyr His Pro Asp Lys Ile Lys Ala Asn His Asn Asp 565 570 575Lys Gln
Glu Ser Ile His Glu Thr Met Ser Gln Ile Asn Glu Ala Tyr 580 585
590Glu Thr Leu Ser Asp Asp Asp Lys Arg Lys Glu Tyr Asp Leu Ser Arg
595 600 605Ser Asn Pro Arg Arg Asn Thr Phe Pro Gln Gly Pro Arg Gln
Asn Asn 610 615 620Met Phe Lys Asn Pro Gly Ser Gly Phe Pro Phe Gly
Asn Gly Phe Lys625 630 635 640Met Asn Phe Gly Leu
64521938DNASaccharomyces cerevisiae 2atgatactga tctcgggata
ctgtctttta gtgtatagcg ttattttgcc agtactgata 60tcggcttcta agttatgtga
tttggctgag ttacaacgat tgaacaagaa tttaaaagta 120gacactgaat
ccttgccaaa ataccaatgg atcgctgggc agttggaaca aaactgcatg
180actgcggatc cagcaagtga aaatatgtca gacgtaattc aactagccaa
tcaaatatac 240tacaaaattg ggctgatcca attatccaac gatcaacatc
taagagctat taacacattt 300gaaaaaatcg tttttaatga aacttacaaa
ggttcttttg ggaagctggc ggaaaagagg 360ctacaagagc tgtatgtcga
ttttgggatg tgggacaagg tgcatcagaa ggatgatcag 420tatgcgaaat
atctgtcctt gaatgaaacc atcagaaaca aaatatcatc caaagacgtt
480tctgtggagg aagatatttc tgagctgcta cgcataacgc cgtacgatgt
taacgtcctc 540tccacgcaca tcgatgttct ttttcacaaa ctagctgaag
aaattgacgt ttcgttagct 600gctgctatca ttttggatta cgaaacaatc
ctcgacaagc atttggctag cttaagcata 660gatacaagac tttcgattca
ttatgtcata tctgttttac agacctttgt acttaactca 720gatgcgtcgt
tcaatataag aaaatgcctt tccattgata tggactatga taaatgtaaa
780aaactaagcc tgactatttc caaattgaac aaggtgaatc catcaaaaag
acagatcctg 840gatccagcaa catatgcatt tgagaacaaa aagtttagaa
gttgggatag aattattgaa 900ttttatttga aggataagaa gccatttatt
acaccaatga aaattcttaa caaagataca 960aactttaaaa acaactactt
ctttttagag gaaattatca aacaattgat agaagacgtt 1020caactgtcga
gacctttggc aaaaaattta ttcgaagatc ccccaataac cgatggtttt
1080gtcaaaccaa aatcatacta tcataccgat tatctagtat acattgattc
cattctttgt 1140caggcttcta gcatgagtcc ggacgtcaag agagctaaac
tggctgcgcc gttctgtaaa 1200aagagtttga ggcattcact aacactagaa
acatggaaac actatcagga tgctaagtcc 1260gagcaaaaac ctttacctga
gacggtattg agtgatgtat ggaattccaa tcctcatttg 1320ctgatgtata
tggtaaactc aatacttaat aaaagtaggt ctaaacctca ttcacagttc
1380aaaaagcaat tatatgacca gataaacaaa tttttccaag ataacggcct
ctcagagtcg 1440accaatccat acgtgatgaa gaacttccga ttattacaga
aacaattaca aacctataaa 1500gagcataaac atcggaattt caaccagcaa
tatttccaac aacaacaaca gcagcaacaa 1560caccaacgac atcaagcacc
cccagcagcg cctaactacg acccaaaaaa ggactattat 1620aaaattcttg
gagtatcgcc tagtgctagt tcgaaagaaa taaggaaagc atatttaaat
1680ttaaccaaaa aataccaccc agacaaaata aaggccaacc ataacgacaa
acaagaatca 1740attcacgaaa ctatgtcaca aatcaatgaa gcgtacgaaa
cattaagtga tgacgataaa 1800aggaaggaat acgatctttc cagatcaaac
ccccgccgca acacttttcc tcaggggcct 1860aggcaaaata acatgttcaa
aaatccagga agtggcttcc cattcggaaa tggctttaaa 1920atgaattttg ggctttga
19383881PRTSaccharomyces cerevisiae 3Met Arg Asn Val Leu Arg Leu
Leu Phe Leu Thr Ala Phe Val Ala Ile1 5 10 15Gly Ser Leu Ala Ala Val
Leu Gly Val Asp Tyr Gly Gln Gln Asn Ile 20 25 30Lys Ala Ile Val Val
Ser Pro Gln Ala Pro Leu Glu Leu Val Leu Thr 35 40 45Pro Glu Ala Lys
Arg Lys Glu Ile Ser Gly Leu Ser Ile Lys Arg Leu 50 55 60Pro Gly Tyr
Gly Lys Asp Asp Pro Asn Gly Ile Glu Arg Ile Tyr Gly65 70 75 80Ser
Ala Val Gly Ser Leu Ala Thr Arg Phe Pro Gln Asn Thr Leu Leu 85 90
95His Leu Lys Pro Leu Leu Gly Lys Ser Leu Glu Asp Glu Thr Thr Val
100 105 110Thr Leu Tyr Ser Lys Gln His Pro Gly Leu Glu Met Val Ser
Thr Asn 115 120 125Arg Ser Thr Ile Ala Phe Leu Val Asp Asn Val Glu
Tyr Pro Leu Glu 130 135 140Glu Leu Val Ala Met Asn Val Gln Glu Ile
Ala Asn Arg Ala Asn Ser145 150 155 160Leu Leu Lys Asp Arg Asp Ala
Arg Thr Glu Asp Phe Val Asn Lys Met 165 170 175Ser Phe Thr Ile Pro
Asp Phe Phe Asp Gln His Gln Arg Lys Ala Leu 180 185 190Leu Asp Ala
Ser Ser Ile Thr Thr Gly Ile Glu Glu Thr Tyr Leu Val 195 200 205Ser
Glu Gly Met Ser Val Ala Val Asn Phe Val Leu Lys Gln Arg Gln 210 215
220Phe Pro Pro Gly Glu Gln Gln His Tyr Ile Val Tyr Asp Met Gly
Ser225 230 235 240Gly Ser Ile Lys Ala Ser Met Phe Ser Ile Leu Gln
Pro Glu Asp Thr 245 250 255Thr Gln Pro Val Thr Ile Glu Phe Glu Gly
Tyr Gly Tyr Asn Pro His 260 265 270Leu Gly Gly Ala Lys Phe Thr Met
Asp Ile Gly Ser Leu Ile Glu Asn 275 280 285Lys Phe Leu Glu Thr His
Pro Ala Ile Arg Thr Asp Glu Leu His Ala 290 295 300Asn Pro Lys Ala
Leu Ala Lys Ile Asn Gln Ala Ala Glu Lys Ala Lys305 310 315 320Leu
Ile Leu Ser Ala Asn Ser Glu Ala Ser Ile Asn Ile Glu Ser Leu 325 330
335Ile Asn Asp Ile Asp Phe Arg Thr Ser Ile Thr Arg Gln Glu Phe Glu
340 345 350Glu Phe Ile Ala Asp Ser Leu Leu Asp Ile Val Lys Pro Ile
Asn Asp 355 360 365Ala Val Thr Lys Gln Phe Gly Gly Tyr Gly Thr Asn
Leu Pro Glu Ile 370 375 380Asn Gly Val Ile Leu Ala Gly Gly Ser Ser
Arg Ile Pro Ile Val Gln385 390 395 400Asp Gln Leu Ile Lys Leu Val
Ser Glu Glu Lys Val Leu Arg Asn Val 405 410 415Asn Ala Asp Glu Ser
Ala Val Asn Gly Val Val Met Arg Gly Ile Lys 420 425 430Leu Ser Asn
Ser Phe Lys Thr Lys Pro Leu Asn Val Val Asp Arg Ser 435 440 445Val
Asn Thr Tyr Ser Phe Lys Leu Ser Asn Glu Ser Glu Leu Tyr Asp 450 455
460Val Phe Thr Arg Gly Ser Ala Tyr Pro Asn Lys Thr Ser Ile Leu
Thr465 470 475 480Asn Thr Thr Asp Ser Ile Pro Asn Asn Phe Thr Ile
Asp Leu Phe Glu 485 490 495Asn Gly Lys Leu Phe Glu Thr Ile Thr Val
Asn Ser Gly Ala Ile Lys 500 505 510Asn Ser Tyr Ser Ser Asp Lys Cys
Ser Ser Gly Val Ala Tyr Asn Ile 515 520 525Thr Phe Asp Leu Ser Ser
Asp Arg Leu Phe Ser Ile Gln Glu Val Asn 530 535 540Cys Ile Cys Gln
Ser Glu Asn Asp Ile Gly Asn Ser Lys Gln Ile Lys545 550 555 560Asn
Lys Gly Ser Arg Leu Ala Phe Thr Ser Glu Asp Val Glu Ile Lys 565 570
575Arg Leu Ser Pro Ser Glu Arg Ser Arg Leu His Glu His Ile Lys Leu
580 585 590Leu Asp Lys Gln Asp Lys Glu Arg Phe Gln Phe Gln Glu Asn
Leu Asn 595 600 605Val Leu Glu Ser Asn Leu Tyr Asp Ala Arg Asn Leu
Leu Met Asp Asp 610 615 620Glu Val Met Gln Asn Gly Pro Lys Ser Gln
Val Glu Glu Leu Ser Glu625 630 635 640Met Val Lys Val Tyr Leu Asp
Trp Leu Glu Asp Ala Ser Phe Asp Thr 645 650 655Asp Pro Glu Asp Ile
Val Ser Arg Ile Arg Glu Ile Gly Ile Leu Lys 660 665 670Lys Lys Ile
Glu Leu Tyr Met Asp Ser Ala Lys Glu Pro Leu Asn Ser 675 680 685Gln
Gln Phe Lys Gly Met Leu Glu Glu Gly His Lys Leu Leu Gln Ala 690 695
700Ile Glu Thr His Lys Asn Thr Val Glu Glu Phe Leu Ser Gln Phe
Glu705 710 715 720Thr Glu Phe Ala Asp Thr Ile Asp Asn Val Arg Glu
Glu Phe Lys Lys 725 730 735Ile Lys Gln Pro Ala Tyr Val Ser Lys Ala
Leu Ser Thr Trp Glu Glu 740 745 750Thr Leu Thr Ser Phe Lys Asn Ser
Ile Ser Glu Ile Glu Lys Phe Leu 755 760 765Ala Lys Asn Leu Phe Gly
Glu Asp Leu Arg Glu His Leu Phe Glu Ile 770 775 780Lys Leu Gln Phe
Asp Met Tyr Arg Thr Lys Leu Glu Glu Lys Leu Arg785 790 795 800Leu
Ile Lys Ser Gly Asp Glu Ser Arg Leu Asn Glu Ile Lys Lys Leu 805 810
815His Leu Arg Asn Phe Arg Leu Gln Lys Arg Lys Glu Glu Lys Leu Lys
820 825 830Arg Lys Leu Glu Gln Glu Lys Ser Arg Asn Asn Asn Glu Thr
Glu Ser 835 840 845Thr Val Ile Asn Ser Ala Asp Asp Lys Thr Thr Ile
Val Asn Asp Lys 850 855 860Thr Thr Glu Ser Asn Pro Ser Ser Glu Glu
Asp Ile Leu His Asp Glu865 870 875 880Leu42646DNASaccharomyces
cerevisiae 4atgcgaaacg ttttaaggct tttattttta acagcttttg ttgctatagg
gtctttagca 60gccgttttag gtgttgatta cggtcagcaa aatatcaagg ccattgtggt
ttctccgcaa 120gccccattag aacttgtgct cacaccagag gcaaaacgga
aggagatatc tggtctttcg 180ataaaaagat taccaggtta tggaaaggat
gatccgaatg ggattgaaag aatctacggt 240tccgctgttg gcagtttagc
aacaaggttt ccccaaaaca cattgttgca tttgaaaccg 300ctacttggga
aatcactaga agatgaaacc actgtaactt tgtattcaaa acaacacccc
360ggtttagaaa tggtatcaac aaatagaagt accatagcct ttttagttga
taatgtggaa 420tatccattgg aagagttagt ggcaatgaat gtccaagaga
ttgccaatag agccaattca 480ctgttgaagg atagagatgc aagaactgag
gactttgtaa acaagatgag ttttacaatt 540cctgactttt ttgaccaaca
tcaaaggaaa gcacttttag atgccagttc aataaccaca 600ggaatcgaag
agacatatct ggttagtgaa gggatgtctg ttgcagttaa ctttgtatta
660aagcagcgcc aatttccacc aggtgaacag cagcattata tcgtatatga
catggggagc 720ggttctatta aggcctcaat gttctctata ttgcagccgg
aggacactac tcagcccgtt 780acaatagaat ttgaaggata tgggtataat
ccacatctag gtggtgcaaa gtttacaatg 840gatattggca gtttgataga
gaataagttt ttggaaacac acccagccat aagaactgat 900gaattgcacg
ctaatcccaa ggccttagca aaaatcaacc aagcagcaga gaaggcaaag
960ttaattttaa gcgccaattc tgaggcaagt attaacatag aatcactgat
caacgatatt 1020gatttccgta cttctataac tagacaggaa ttcgaagaat
ttattgcaga ctcgttattg 1080gacattgtca aacccataaa tgacgctgtt
acaaaacaat tcggtggcta tggaacaaat 1140ttacctgaga taaatggggt
cattttggcg ggaggctctt cccgaattcc cattgtgcag 1200gatcaattaa
tcaaactcgt atccgaagaa aaagtgttga gaaatgtcaa tgctgatgaa
1260tcagctgtga atggtgttgt tatgagaggg atcaagttat ctaattcgtt
taagaccaag 1320ccgttaaatg ttgttgaccg ttctgtaaat acttattcat
tcaaattatc aaacgaatct 1380gaactgtatg atgtgttcac gcgcggaagt
gcttatccaa acaaaacatc tattttgaca 1440aacacgactg attcgattcc
taataatttt accattgact tatttgagaa tggtaaattg 1500ttcgaaacta
tcacagttaa ttcaggagct ataaagaatt catattcctc tgataagtgc
1560tcgtcaggag ttgcgtataa cattactttc gacttgtcca gtgatagatt
attctctatt 1620caagaggtta actgcatttg tcagagcgaa aatgacatag
gtaactccaa gcaaattaag 1680aacaaaggca gccgtttggc ttttacttct
gaggatgttg agatcaaaag gctttctcct 1740tcagaacgtt cgcgtttgca
tgagcatatc aagttgctcg ataaacagga taaggaaaga 1800tttcaattcc
aagaaaattt aaacgttctt gaaagtaact tgtatgatgc tagaaacctg
1860ctaatggatg atgaagttat gcaaaatgga ccaaaatccc aagtagaaga
gttatcggag 1920atggttaaag tatatttgga ttggctcgaa gatgcatcct
ttgatactga ccctgaggat 1980atagttagca gaattagaga aattggaata
ttaaaaaaga aaatagaact ttacatggat 2040tctgcaaagg aacctttgaa
ctctcaacaa tttaaaggaa tgcttgaaga aggccataag 2100ttacttcagg
ctatagaaac ccataagaat accgttgaag aatttttgag tcaatttgaa
2160accgagtttg cggataccat agataatgtt agagaagaat ttaaaaagat
taagcaacca 2220gcgtatgtgt cgaaggcgtt atctacatgg gaggaaacct
taacctcttt taaaaattcc 2280attagcgaaa tagagaagtt cctggcaaaa
aacctatttg gcgaagacct tcgtgaacat 2340ttatttgaaa tcaaattaca
atttgatatg tatcgtacga aactagagga aaaactgcgt 2400ttaataaaaa
gcggtgatga aagtcgctta aatgaaataa agaagttaca tttaagaaac
2460ttccgcctac aaaagagaaa ggaggaaaag ttgaaaagaa agcttgaaca
ggaaaaaagc 2520agaaacaaca atgaaacaga atcgacagta atcaactcgg
ctgacgataa aactactatt 2580gtcaatgaca agaccaccga gtcgaatcca
agttctgagg aagacatttt gcatgatgaa 2640ttatag
26465377PRTSaccharomyces cerevisiae 5Met Ile Pro Lys Leu Tyr Ile
His Leu Ile Leu Ser Leu Leu Leu Leu1 5 10 15Pro Leu Ile Leu Ala Gln
Asp Tyr Tyr Ala Ile Leu Glu Ile Asp Lys 20 25 30Asp Ala Thr Glu Lys
Glu Ile Lys Ser Ala Tyr Arg Gln Leu Ser Lys 35 40 45Lys Tyr His Pro
Asp Lys Asn Ala Gly Ser Glu Glu Ala His Gln Lys 50 55 60Phe Ile Glu
Val Gly Glu Ala Tyr Asp Val Leu Ser Asp Pro Glu Lys65 70 75 80Lys
Lys Ile Tyr Asp Gln Phe Gly Ala Asp Ala Val Lys Asn Gly Gly 85 90
95Gly Gly Gly Gly Pro Gly Gly Pro Gly Ala Gly Gly Phe His Asp Pro
100 105 110Phe Asp Ile Phe Glu Arg Met Phe Gln Gly Gly His Gly Gly
Pro Gly 115 120 125Gly Gly Phe Gly Gln Arg Gln Arg Gln Arg Gly Pro
Met Ile Lys Val 130 135 140Gln Glu Lys Leu Ser Leu Lys Gln Phe Tyr
Ser Gly Ser Ser Ile Glu145 150
155 160Phe Thr Leu Asn Leu Asn Asp Glu Cys Asp Ala Cys His Gly Ser
Gly 165 170 175Ser Ala Asp Gly Lys Leu Ala Gln Cys Pro Asp Cys Gln
Gly Arg Gly 180 185 190Val Ile Ile Gln Val Leu Arg Met Gly Ile Met
Thr Gln Gln Ile Gln 195 200 205Gln Met Cys Gly Arg Cys Gly Gly Thr
Gly Gln Ile Ile Lys Asn Glu 210 215 220Cys Lys Thr Cys His Gly Lys
Lys Val Thr Lys Lys Asn Lys Phe Phe225 230 235 240His Val Asp Val
Pro Pro Gly Ala Pro Arg Asn Tyr Met Asp Thr Arg 245 250 255Val Gly
Glu Ala Glu Lys Gly Pro Asp Phe Asp Ala Gly Asp Leu Val 260 265
270Ile Glu Phe Lys Glu Lys Asp Thr Glu Asn Met Gly Tyr Arg Arg Arg
275 280 285Gly Asp Asn Leu Tyr Arg Thr Glu Val Leu Ser Ala Ala Glu
Ala Leu 290 295 300Tyr Gly Gly Trp Gln Arg Thr Ile Glu Phe Leu Asp
Glu Asn Lys Pro305 310 315 320Val Lys Leu Ser Arg Pro Ala His Val
Val Val Ser Asn Gly Glu Val 325 330 335Glu Val Val Lys Gly Phe Gly
Met Pro Lys Gly Ser Lys Gly Tyr Gly 340 345 350Asp Leu Tyr Ile Asp
Tyr Val Val Val Met Pro Lys Thr Phe Lys Ser 355 360 365Gly Gln Asn
Met Leu Lys Asp Glu Leu 370 37561134DNASaccharomyces cerevisiae
6atgattccaa aattatatat acatttgata ctatctttat tgttgttgcc gctaattttg
60gcgcaggatt attatgcaat actagagata gacaaagatg ccactgagaa ggaaatcaaa
120tcagcgtaca gacaattgtc taagaagtac catccggata aaaatgctgg
gagcgaagaa 180gcccatcaaa aattcattga agtcggcgag gcatacgatg
tattgagcga tcctgaaaag 240aaaaagattt atgaccagtt tggtgcagat
gctgtaaaga atggcggtgg cggtggcggt 300ccaggaggcc ctggcgcagg
tggattccac gatccgtttg acatattcga acggatgttt 360caaggaggtc
atggaggtcc tggcggcgga tttggccaga gacagaggca gcgtggtcca
420atgatcaagg tccaggaaaa actatcttta aagcagtttt attccgggtc
ctcgatagaa 480tttactttaa acctaaacga tgaatgtgat gcatgccatg
gtagtggctc tgcagatggt 540aagctggccc aatgtcccga ttgtcaaggt
cgtggggtta taatacaagt gctgcgcatg 600ggtattatga cgcagcagat
tcaacagatg tgtggtaggt gtggtggtac gggacaaatt 660atcaaaaatg
aatgcaaaac atgtcacggc aaaaaagtta ccaaaaagaa caagttcttc
720cacgttgacg ttccaccagg cgcaccaaga aactacatgg acacaagagt
cggcgaggct 780gaaaaagggc ctgactttga cgccggtgac ttggtcatag
aattcaagga aaaggatact 840gagaacatgg gttacagaag aagaggcgac
aatctgtaca gaacagaagt tctttctgct 900gcggaagcgc tatacggcgg
atggcaaaga acgatagaat tccttgatga gaacaagccc 960gttaagttat
ctagacccgc tcatgtagtt gtctccaatg gcgaagttga agtcgtgaag
1020ggattcggca tgcccaaggg tagcaagggt tacggtgatt tgtacataga
ctacgtcgtt 1080gtcatgccaa agactttcaa atctgggcaa aatatgctca
aagatgagtt gtag 11347682PRTSaccharomyces cerevisiae 7Met Phe Phe
Asn Arg Leu Ser Ala Gly Lys Leu Leu Val Pro Leu Ser1 5 10 15Val Val
Leu Tyr Ala Leu Phe Val Val Ile Leu Pro Leu Gln Asn Ser 20 25 30Phe
His Ser Ser Asn Val Leu Val Arg Gly Ala Asp Asp Val Glu Asn 35 40
45Tyr Gly Thr Val Ile Gly Ile Asp Leu Gly Thr Thr Tyr Ser Cys Val
50 55 60Ala Val Met Lys Asn Gly Lys Thr Glu Ile Leu Ala Asn Glu Gln
Gly65 70 75 80Asn Arg Ile Thr Pro Ser Tyr Val Ala Phe Thr Asp Asp
Glu Arg Leu 85 90 95Ile Gly Asp Ala Ala Lys Asn Gln Val Ala Ala Asn
Pro Gln Asn Thr 100 105 110Ile Phe Asp Ile Lys Arg Leu Ile Gly Leu
Lys Tyr Asn Asp Arg Ser 115 120 125Val Gln Lys Asp Ile Lys His Leu
Pro Phe Asn Val Val Asn Lys Asp 130 135 140Gly Lys Pro Ala Val Glu
Val Ser Val Lys Gly Glu Lys Lys Val Phe145 150 155 160Thr Pro Glu
Glu Ile Ser Gly Met Ile Leu Gly Lys Met Lys Gln Ile 165 170 175Ala
Glu Asp Tyr Leu Gly Thr Lys Val Thr His Ala Val Val Thr Val 180 185
190Pro Ala Tyr Phe Asn Asp Ala Gln Arg Gln Ala Thr Lys Asp Ala Gly
195 200 205Thr Ile Ala Gly Leu Asn Val Leu Arg Ile Val Asn Glu Pro
Thr Ala 210 215 220Ala Ala Ile Ala Tyr Gly Leu Asp Lys Ser Asp Lys
Glu His Gln Ile225 230 235 240Ile Val Tyr Asp Leu Gly Gly Gly Thr
Phe Asp Val Ser Leu Leu Ser 245 250 255Ile Glu Asn Gly Val Phe Glu
Val Gln Ala Thr Ser Gly Asp Thr His 260 265 270Leu Gly Gly Glu Asp
Phe Asp Tyr Lys Ile Val Arg Gln Leu Ile Lys 275 280 285Ala Phe Lys
Lys Lys His Gly Ile Asp Val Ser Asp Asn Asn Lys Ala 290 295 300Leu
Ala Lys Leu Lys Arg Glu Ala Glu Lys Ala Lys Arg Ala Leu Ser305 310
315 320Ser Gln Met Ser Thr Arg Ile Glu Ile Asp Ser Phe Val Asp Gly
Ile 325 330 335Asp Leu Ser Glu Thr Leu Thr Arg Ala Lys Phe Glu Glu
Leu Asn Leu 340 345 350Asp Leu Phe Lys Lys Thr Leu Lys Pro Val Glu
Lys Val Leu Gln Asp 355 360 365Ser Gly Leu Glu Lys Lys Asp Val Asp
Asp Ile Val Leu Val Gly Gly 370 375 380Ser Thr Arg Ile Pro Lys Val
Gln Gln Leu Leu Glu Ser Tyr Phe Asp385 390 395 400Gly Lys Lys Ala
Ser Lys Gly Ile Asn Pro Asp Glu Ala Val Ala Tyr 405 410 415Gly Ala
Ala Val Gln Ala Gly Val Leu Ser Gly Glu Glu Gly Val Glu 420 425
430Asp Ile Val Leu Leu Asp Val Asn Ala Leu Thr Leu Gly Ile Glu Thr
435 440 445Thr Gly Gly Val Met Thr Pro Leu Ile Lys Arg Asn Thr Ala
Ile Pro 450 455 460Thr Lys Lys Ser Gln Ile Phe Ser Thr Ala Val Asp
Asn Gln Pro Thr465 470 475 480Val Met Ile Lys Val Tyr Glu Gly Glu
Arg Ala Met Ser Lys Asp Asn 485 490 495Asn Leu Leu Gly Lys Phe Glu
Leu Thr Gly Ile Pro Pro Ala Pro Arg 500 505 510Gly Val Pro Gln Ile
Glu Val Thr Phe Ala Leu Asp Ala Asn Gly Ile 515 520 525Leu Lys Val
Ser Ala Thr Asp Lys Gly Thr Gly Lys Ser Glu Ser Ile 530 535 540Thr
Ile Thr Asn Asp Lys Gly Arg Leu Thr Gln Glu Glu Ile Asp Arg545 550
555 560Met Val Glu Glu Ala Glu Lys Phe Ala Ser Glu Asp Ala Ser Ile
Lys 565 570 575Ala Lys Val Glu Ser Arg Asn Lys Leu Glu Asn Tyr Ala
His Ser Leu 580 585 590Lys Asn Gln Val Asn Gly Asp Leu Gly Glu Lys
Leu Glu Glu Glu Asp 595 600 605Lys Glu Thr Leu Leu Asp Ala Ala Asn
Asp Val Leu Glu Trp Leu Asp 610 615 620Asp Asn Phe Glu Thr Ala Ile
Ala Glu Asp Phe Asp Glu Lys Phe Glu625 630 635 640Ser Leu Ser Lys
Val Ala Tyr Pro Ile Thr Ser Lys Leu Tyr Gly Gly 645 650 655Ala Asp
Gly Ser Gly Ala Ala Asp Tyr Asp Asp Glu Asp Glu Asp Asp 660 665
670Asp Gly Asp Tyr Phe Glu His Asp Glu Leu 675
68082049DNASaccharomyces cerevisiae 8atgtttttca acagactaag
cgctggcaag ctgctggtac cactctccgt ggtcctgtac 60gcccttttcg tggtaatatt
acctttacag aattctttcc actcctccaa tgttttagtt 120agaggtgccg
atgatgtaga aaactacgga actgttatcg gtattgactt aggtactact
180tattcctgtg ttgctgtgat gaaaaatggt aagactgaaa ttcttgctaa
tgagcaaggt 240aacagaatca ccccatctta cgtggcattc accgatgatg
aaagattgat tggtgatgct 300gcaaagaacc aagttgctgc caatcctcaa
aacaccatct tcgacattaa gagattgatc 360ggtttgaaat ataacgacag
atctgttcag aaggatatca agcacttgcc atttaatgtg 420gttaataaag
atgggaagcc cgctgtagaa gtaagtgtca aaggagaaaa gaaggttttt
480actccagaag aaatttctgg tatgatcttg ggtaagatga aacaaattgc
cgaagattat 540ttaggcacta aggttaccca tgctgtcgtt actgttcctg
cttatttcaa tgacgcgcaa 600agacaagcca ccaaggatgc tggtaccatc
gctggtttga acgttttgag aattgttaat 660gaaccaaccg cagccgccat
tgcctacggt ttggataaat ctgataagga acatcaaatt 720attgtttatg
atttgggtgg tggtactttc gatgtctctc tattgtctat tgaaaacggt
780gttttcgaag tccaagccac ttctggtgat actcatttag gtggtgaaga
ttttgactat 840aagatcgttc gtcaattgat aaaagctttc aagaagaagc
atggtattga tgtgtctgac 900aacaacaagg ccctagctaa attgaagaga
gaagctgaaa aggctaaacg tgccttgtcc 960agccaaatgt ccacccgtat
tgaaattgac tccttcgttg atggtatcga cttaagtgaa 1020accttgacca
gagctaagtt tgaggaatta aacctagatc tattcaagaa gaccttgaag
1080cctgtcgaga aggttttgca agattctggt ttggaaaaga aggatgttga
tgatatcgtt 1140ttggttggtg gttctactag aattccaaag gtccaacaat
tgttagaatc atactttgat 1200ggtaagaagg cctccaaggg tattaaccca
gatgaagctg ttgcatacgg tgcagccgtt 1260caagctggtg tcttatccgg
tgaagaaggt gtcgaagata ttgttttatt ggatgtcaac 1320gctttgactc
ttggtattga aaccactggt ggtgtcatga ctccattaat taagagaaat
1380actgctattc ctacaaagaa atcccaaatt ttctctactg ccgttgacaa
ccaaccaacc 1440gttatgatca aggtatacga gggtgaaaga gccatgtcta
aggacaacaa tctattaggt 1500aagtttgaat taaccggcat tccaccagca
ccaagaggtg tacctcaaat tgaagtcaca 1560tttgcacttg acgctaatgg
tattctgaag gtgtctgcca cagataaggg aactggtaaa 1620tccgaatcta
tcaccatcac taacgataaa ggtagattaa cccaagaaga gattgataga
1680atggttgaag aggctgaaaa attcgcttct gaagacgctt ctatcaaggc
caaggttgaa 1740tctagaaaca aattagaaaa ctacgctcac tctttgaaaa
accaagttaa tggtgaccta 1800ggtgaaaaat tggaagaaga agacaaggaa
accttattag atgctgctaa cgatgtttta 1860gaatggttag atgataactt
tgaaaccgcc attgctgaag actttgatga aaagttcgaa 1920tctttgtcca
aggtcgctta tccaattact tctaagttgt acggaggtgc tgatggttct
1980ggtgccgctg attatgacga cgaagatgaa gatgacgatg gtgattattt
cgaacacgac 2040gaattgtag 20499421PRTSaccharomyces cerevisiae 9Met
Val Arg Ile Leu Pro Ile Ile Leu Ser Ala Leu Ser Ser Lys Leu1 5 10
15Val Ala Ser Thr Ile Leu His Ser Ser Ile His Ser Val Pro Ser Gly
20 25 30Gly Glu Ile Ile Ser Ala Glu Asp Leu Lys Glu Leu Glu Ile Ser
Gly 35 40 45Asn Ser Ile Cys Val Asp Asn Arg Cys Tyr Pro Lys Ile Phe
Glu Pro 50 55 60Arg His Asp Trp Gln Pro Ile Leu Pro Gly Gln Glu Leu
Pro Gly Gly65 70 75 80Leu Asp Ile Arg Ile Asn Met Asp Thr Gly Leu
Lys Glu Ala Lys Leu 85 90 95Asn Asp Glu Lys Asn Val Gly Asp Asn Gly
Ser His Glu Leu Ile Val 100 105 110Ser Ser Glu Asp Met Lys Ala Ser
Pro Gly Asp Tyr Glu Phe Ser Ser 115 120 125Asp Phe Lys Glu Met Arg
Asn Ile Ile Asp Ser Asn Pro Thr Leu Ser 130 135 140Ser Gln Asp Ile
Ala Arg Leu Glu Asp Ser Phe Asp Arg Ile Met Glu145 150 155 160Phe
Ala His Asp Tyr Lys His Gly Tyr Lys Ile Ile Thr His Glu Phe 165 170
175Ala Leu Leu Ala Asn Leu Ser Leu Asn Glu Asn Leu Pro Leu Thr Leu
180 185 190Arg Glu Leu Ser Thr Arg Val Ile Thr Ser Cys Leu Arg Asn
Asn Pro 195 200 205Pro Val Val Glu Phe Ile Asn Glu Ser Phe Pro Asn
Phe Lys Ser Lys 210 215 220Ile Met Ala Ala Leu Ser Asn Leu Asn Asp
Ser Asn His Arg Ser Ser225 230 235 240Asn Ile Leu Ile Lys Arg Tyr
Leu Ser Ile Leu Asn Glu Leu Pro Val 245 250 255Thr Ser Glu Asp Leu
Pro Ile Tyr Ser Thr Val Val Leu Gln Asn Val 260 265 270Tyr Glu Arg
Asn Asn Lys Asp Lys Gln Leu Gln Ile Lys Val Leu Glu 275 280 285Leu
Ile Ser Lys Ile Leu Lys Ala Asp Met Tyr Glu Asn Asp Asp Thr 290 295
300Asn Leu Ile Leu Phe Lys Arg Asn Ala Glu Asn Trp Ser Ser Asn
Leu305 310 315 320Gln Glu Trp Ala Asn Glu Phe Gln Glu Met Val Gln
Asn Lys Ser Ile 325 330 335Asp Glu Leu His Thr Arg Thr Phe Phe Asp
Thr Leu Tyr Asn Leu Lys 340 345 350Lys Ile Phe Lys Ser Asp Ile Thr
Ile Asn Lys Gly Phe Leu Asn Trp 355 360 365Leu Ala Gln Gln Cys Lys
Ala Arg Gln Ser Asn Leu Asp Asn Gly Leu 370 375 380Gln Glu Arg Asp
Thr Glu Gln Asp Ser Phe Asp Lys Lys Leu Ile Asp385 390 395 400Ser
Arg His Leu Ile Phe Gly Asn Pro Met Ala His Arg Ile Lys Asn 405 410
415Phe Arg Asp Glu Leu 420101266DNASaccharomyces cerevisiae
10atggtccgga ttcttcccat aattttgagc gccctatctt cgaaattagt ggcgagtaca
60atattgcatt catccataca ctcagtgcca tctggaggcg aaatcatatc tgcagaagat
120cttaaagaac ttgaaatttc agggaattcg atctgcgttg ataatcgttg
ctatcctaag 180atatttgaac caagacacga ttggcagccc atactgccag
gtcaagaact ccccggtggt 240ttggacatta gaataaacat ggacacaggt
ttaaaagagg caaaactaaa tgatgagaag 300aatgtcggtg ataatggtag
ccatgagtta attgtatctt cagaagacat gaaagcatcg 360cctggtgact
atgaattttc cagtgatttc aaagaaatga gaaacatcat agattctaac
420ccgactttat cttcacagga cattgccaga ttggaggata gttttgatag
aataatggaa 480tttgcgcatg attacaagca cggctacaaa attattaccc
atgaattcgc cctcttggcc 540aaccttagtc tcaatgaaaa tttgccgtta
acattgagag agctcagtac tagagtcatt 600accagctgct tgagaaacaa
tcctcctgta gtcgagttca ttaatgaaag ttttccaaat 660tttaaaagca
aaatcatggc cgctctgtca aatttgaatg attctaacca cagatcctct
720aatatcctaa taaaaagata cttgtccatt ttaaacgaat tacctgtcac
atccgaagat 780cttcctatat actctacggt tgttttacaa aatgtatatg
aaagaaacaa caaggacaaa 840cagttacaaa taaaagtcct ggagttgatc
agcaaaattt tgaaggccga catgtacgaa 900aatgacgata caaatctaat
tttgttcaaa agaaatgctg agaattggtc gtcaaatctg 960caagagtggg
caaacgagtt ccaagagatg gtccagaaca aaagtataga tgaactacat
1020acaagaacgt tttttgacac cctttacaac ttgaagaaaa ttttcaaaag
tgacatcacg 1080atcaacaaag ggtttttgaa ttggttagcg caacaatgta
aagccaggca atctaacttg 1140gacaatgggc tccaagagag agatactgaa
caagactcat ttgataagaa acttatcgac 1200agcagacact tgatctttgg
caaccccatg gctcatagaa taaaaaattt cagagatgaa 1260ctctga
126611135PRTSaccharomyces cerevisiae 11Met Met Phe Asn Ile Tyr Leu
Phe Val Thr Phe Phe Ser Thr Ile Leu1 5 10 15Ala Gly Ser Leu Ser Asp
Leu Glu Ile Gly Ile Ile Lys Arg Ile Pro 20 25 30Val Glu Asp Cys Leu
Ile Lys Ala Met Pro Gly Asp Lys Val Lys Val 35 40 45His Tyr Thr Gly
Ser Leu Leu Glu Ser Gly Thr Val Phe Asp Ser Ser 50 55 60Tyr Ser Arg
Gly Ser Pro Ile Ala Phe Glu Leu Gly Val Gly Arg Val65 70 75 80Ile
Lys Gly Trp Asp Gln Gly Val Ala Gly Met Cys Val Gly Glu Lys 85 90
95Arg Lys Leu Gln Ile Pro Ser Ser Leu Ala Tyr Gly Glu Arg Gly Val
100 105 110Pro Gly Val Ile Pro Pro Ser Ala Asp Leu Val Phe Asp Val
Glu Leu 115 120 125Val Asp Val Lys Ser Ala Ala 130
13512408DNASaccharomyces cerevisiae 12atgatgttta atatttacct
tttcgtcact tttttttcca ccattcttgc aggttccctg 60tcagatttgg aaatcggtat
tatcaagaga ataccggtag aagattgctt aattaaggca 120atgccaggtg
ataaagttaa ggttcattat acaggatctt tattagaatc gggaactgta
180tttgactcaa gttattcaag aggctctcct atcgcttttg aacttggcgt
tggcagagta 240attaaaggtt gggatcaagg tgttgccggc atgtgcgttg
gcgaaaaaag aaagctgcaa 300attccaagtt ctttggccta cggagaaaga
ggtgtcccag gcgtcattcc tccaagtgct 360gatttggtgt ttgatgtcga
attggtagac gtgaaatcag ccgcctag 40813642PRTSaccharomyces cerevisiae
13Met Ser Lys Ala Val Gly Ile Asp Leu Gly Thr Thr Tyr Ser Cys Val1
5 10 15Ala His Phe Ala Asn Asp Arg Val Asp Ile Ile Ala Asn Asp Gln
Gly 20 25 30Asn Arg Thr Thr Pro Ser Phe Val Ala Phe Thr Asp Thr Glu
Arg Leu 35 40 45Ile Gly Asp Ala Ala Lys Asn Gln Ala Ala Met Asn Pro
Ser Asn Thr 50 55 60Val Phe Asp Ala Lys Arg Leu Ile Gly Arg Asn Phe
Asn Asp Pro Glu65 70 75 80Val Gln Ala Asp Met Lys His Phe Pro Phe
Lys Leu Ile Asp Val Asp 85 90 95Gly Lys Pro Gln Ile Gln Val Glu Phe
Lys Gly Glu Thr Lys Asn Phe 100 105 110Thr Pro Glu Gln Ile Ser Ser
Met Val Leu Gly Lys Met Lys Glu Thr 115 120 125Ala Glu Ser Tyr Leu
Gly Ala Lys Val Asn Asp Ala Val Val Thr Val 130 135 140Pro Ala Tyr
Phe Asn Asp Ser Gln Arg Gln Ala Thr Lys Asp Ala Gly145 150
155 160Thr Ile Ala Gly Leu Asn Val Leu Arg Ile Ile Asn Glu Pro Thr
Ala 165 170 175Ala Ala Ile Ala Tyr Gly Leu Asp Lys Lys Gly Lys Glu
Glu His Val 180 185 190Leu Ile Phe Asp Leu Gly Gly Gly Thr Phe Asp
Val Ser Leu Leu Phe 195 200 205Ile Glu Asp Gly Ile Phe Glu Val Lys
Ala Thr Ala Gly Asp Thr His 210 215 220Leu Gly Gly Glu Asp Phe Asp
Asn Arg Leu Val Asn His Phe Ile Gln225 230 235 240Glu Phe Lys Arg
Lys Asn Lys Lys Asp Leu Ser Thr Asn Gln Arg Ala 245 250 255Leu Arg
Arg Leu Arg Thr Ala Cys Glu Arg Ala Lys Arg Thr Leu Ser 260 265
270Ser Ser Ala Gln Thr Ser Val Glu Ile Asp Ser Leu Phe Glu Gly Ile
275 280 285Asp Phe Tyr Thr Ser Ile Thr Arg Ala Arg Phe Glu Glu Leu
Cys Ala 290 295 300Asp Leu Phe Arg Ser Thr Leu Asp Pro Val Glu Lys
Val Leu Arg Asp305 310 315 320Ala Lys Leu Asp Lys Ser Gln Val Asp
Glu Ile Val Leu Val Gly Gly 325 330 335Ser Thr Arg Ile Pro Lys Val
Gln Lys Leu Val Thr Asp Tyr Phe Asn 340 345 350Gly Lys Glu Pro Asn
Arg Ser Ile Asn Pro Asp Glu Ala Val Ala Tyr 355 360 365Gly Ala Ala
Val Gln Ala Ala Ile Leu Thr Gly Asp Glu Ser Ser Lys 370 375 380Thr
Gln Asp Leu Leu Leu Leu Asp Val Ala Pro Leu Ser Leu Gly Ile385 390
395 400Glu Thr Ala Gly Gly Val Met Thr Lys Leu Ile Pro Arg Asn Ser
Thr 405 410 415Ile Ser Thr Lys Lys Phe Glu Ile Phe Ser Thr Tyr Ala
Asp Asn Gln 420 425 430Pro Gly Val Leu Ile Gln Val Phe Glu Gly Glu
Arg Ala Lys Thr Lys 435 440 445Asp Asn Asn Leu Leu Gly Lys Phe Glu
Leu Ser Gly Ile Pro Pro Ala 450 455 460Pro Arg Gly Val Pro Gln Ile
Glu Val Thr Phe Asp Val Asp Ser Asn465 470 475 480Gly Ile Leu Asn
Val Ser Ala Val Glu Lys Gly Thr Gly Lys Ser Asn 485 490 495Lys Ile
Thr Ile Thr Asn Asp Lys Gly Arg Leu Ser Lys Glu Asp Ile 500 505
510Glu Lys Met Val Ala Glu Ala Glu Lys Phe Lys Glu Glu Asp Glu Lys
515 520 525Glu Ser Gln Arg Ile Ala Ser Lys Asn Gln Leu Glu Ser Ile
Ala Tyr 530 535 540Ser Leu Lys Asn Thr Ile Ser Glu Ala Gly Asp Lys
Leu Glu Gln Ala545 550 555 560Asp Lys Asp Thr Val Thr Lys Lys Ala
Glu Glu Thr Ile Ser Trp Leu 565 570 575Asp Ser Asn Thr Thr Ala Ser
Lys Glu Glu Phe Asp Asp Lys Leu Lys 580 585 590Glu Leu Gln Asp Ile
Ala Asn Pro Ile Met Ser Lys Leu Tyr Gln Ala 595 600 605Gly Gly Ala
Pro Gly Gly Ala Ala Gly Gly Ala Pro Gly Gly Phe Pro 610 615 620Gly
Gly Ala Pro Pro Ala Pro Glu Ala Glu Gly Pro Thr Val Glu Glu625 630
635 640Val Asp141929DNASaccharomyces cerevisiae 14atgtcaaaag
ctgtcggtat tgatttaggt acaacatact cgtgtgttgc tcactttgct 60aatgatcgtg
tggacattat tgccaacgat caaggtaaca gaaccactcc atcttttgtc
120gctttcactg acactgaaag attgattggt gatgctgcta agaatcaagc
tgctatgaat 180ccttcgaata ccgttttcga cgctaagcgt ttgatcggta
gaaacttcaa cgacccagaa 240gtgcaggctg acatgaagca cttcccattc
aagttgatcg atgttgacgg taagcctcaa 300attcaagttg aatttaaggg
tgaaaccaag aactttaccc cagaacaaat ctcctccatg 360gtcttgggta
agatgaagga aactgccgaa tcttacttgg gagccaaggt caatgacgct
420gtcgtcactg tcccagctta cttcaacgat tctcaaagac aagctaccaa
ggatgctggt 480accattgctg gtttgaatgt cttgcgtatt attaacgaac
ctaccgccgc tgccattgct 540tacggtttgg acaagaaggg taaggaagaa
cacgtcttga ttttcgactt gggtggtggt 600actttcgatg tctctttgtt
gttcattgaa gacggtatct ttgaagttaa ggccaccgct 660ggtgacaccc
atttgggtgg tgaagatttt gacaacagat tggtcaacca cttcatccaa
720gaattcaaga gaaagaacaa gaaggacttg tctaccaacc aaagagcttt
gagaagatta 780agaaccgctt gtgaaagagc caagagaact ttgtcttcct
ccgctcaaac ttccgttgaa 840attgactctt tgttcgaagg tatcgatttc
tacacttcca tcaccagagc cagattcgaa 900gaattgtgtg ctgacttgtt
cagatctact ttggacccag ttgaaaaggt cttgagagat 960gctaaattgg
acaaatctca agtcgatgaa attgtcttgg tcggtggttc taccagaatt
1020ccaaaggtcc aaaaattggt cactgactac ttcaacggta aggaaccaaa
cagatctatc 1080aacccagatg aagctgttgc ttacggtgct gctgttcaag
ctgctatttt gactggtgac 1140gaatcttcca agactcaaga tctattgttg
ttggatgtcg ctccattatc cttgggtatt 1200gaaactgctg gtggtgtcat
gaccaagttg attccaagaa actctaccat ttcaacaaag 1260aagttcgaga
tcttttccac ttatgctgat aaccaaccag gtgtcttgat tcaagtcttt
1320gaaggtgaaa gagccaagac taaggacaac aacttgttgg gtaagttcga
attgagtggt 1380attccaccag ctccaagagg tgtcccacaa attgaagtca
ctttcgatgt cgactctaac 1440ggtattttga atgtttccgc cgtcgaaaag
ggtactggta agtctaacaa gatcactatt 1500accaacgaca agggtagatt
gtccaaggaa gatatcgaaa agatggttgc tgaagccgaa 1560aaattcaagg
aagaagatga aaaggaatct caaagaattg cttccaagaa ccaattggaa
1620tccattgctt actctttgaa gaacaccatt tctgaagctg gtgacaaatt
ggaacaagct 1680gacaaggaca ccgtcaccaa gaaggctgaa gagactattt
cttggttaga cagcaacacc 1740actgccagca aggaagaatt cgatgacaag
ttgaaggagt tgcaagacat tgccaaccca 1800atcatgtcta agttgtacca
agctggtggt gctccaggtg gcgctgcagg tggtgctcca 1860ggcggtttcc
caggtggtgc tcctccagct ccagaggctg aaggtccaac cgttgaagaa
1920gttgattaa 192915639PRTSaccharomyces cerevisiae 15Met Ser Lys
Ala Val Gly Ile Asp Leu Gly Thr Thr Tyr Ser Cys Val1 5 10 15Ala His
Phe Ser Asn Asp Arg Val Asp Ile Ile Ala Asn Asp Gln Gly 20 25 30Asn
Arg Thr Thr Pro Ser Phe Val Gly Phe Thr Asp Thr Glu Arg Leu 35 40
45Ile Gly Asp Ala Ala Lys Asn Gln Ala Ala Met Asn Pro Ala Asn Thr
50 55 60Val Phe Asp Ala Lys Arg Leu Ile Gly Arg Asn Phe Asn Asp Pro
Glu65 70 75 80Val Gln Gly Asp Met Lys His Phe Pro Phe Lys Leu Ile
Asp Val Asp 85 90 95Gly Lys Pro Gln Ile Gln Val Glu Phe Lys Gly Glu
Thr Lys Asn Phe 100 105 110Thr Pro Glu Gln Ile Ser Ser Met Val Leu
Gly Lys Met Lys Glu Thr 115 120 125Ala Glu Ser Tyr Leu Gly Ala Lys
Val Asn Asp Ala Val Val Thr Val 130 135 140Pro Ala Tyr Phe Asn Asp
Ser Gln Arg Gln Ala Thr Lys Asp Ala Gly145 150 155 160Thr Ile Ala
Gly Leu Asn Val Leu Arg Ile Ile Asn Glu Pro Thr Ala 165 170 175Ala
Ala Ile Ala Tyr Gly Leu Asp Lys Lys Gly Lys Glu Glu His Val 180 185
190Leu Ile Phe Asp Leu Gly Gly Gly Thr Phe Asp Val Ser Leu Leu Ser
195 200 205Ile Glu Asp Gly Ile Phe Glu Val Lys Ala Thr Ala Gly Asp
Thr His 210 215 220Leu Gly Gly Glu Asp Phe Asp Asn Arg Leu Val Asn
His Phe Ile Gln225 230 235 240Glu Phe Lys Arg Lys Asn Lys Lys Asp
Leu Ser Thr Asn Gln Arg Ala 245 250 255Leu Arg Arg Leu Arg Thr Ala
Cys Glu Arg Ala Lys Arg Thr Leu Ser 260 265 270Ser Ser Ala Gln Thr
Ser Val Glu Ile Asp Ser Leu Phe Glu Gly Ile 275 280 285Asp Phe Tyr
Thr Ser Ile Thr Arg Ala Arg Phe Glu Glu Leu Cys Ala 290 295 300Asp
Leu Phe Arg Ser Thr Leu Asp Pro Val Glu Lys Val Leu Arg Asp305 310
315 320Ala Lys Leu Asp Lys Ser Gln Val Asp Glu Ile Val Leu Val Gly
Gly 325 330 335Ser Thr Arg Ile Pro Lys Val Gln Lys Leu Val Thr Asp
Tyr Phe Asn 340 345 350Gly Lys Glu Pro Asn Arg Ser Ile Asn Pro Asp
Glu Ala Val Ala Tyr 355 360 365Gly Ala Ala Val Gln Ala Ala Ile Leu
Thr Gly Asp Glu Ser Ser Lys 370 375 380Thr Gln Asp Leu Leu Leu Leu
Asp Val Ala Pro Leu Ser Leu Gly Ile385 390 395 400Glu Thr Ala Gly
Gly Val Met Thr Lys Leu Ile Pro Arg Asn Ser Thr 405 410 415Ile Pro
Thr Lys Lys Ser Glu Val Phe Ser Thr Tyr Ala Asp Asn Gln 420 425
430Pro Gly Val Leu Ile Gln Val Phe Glu Gly Glu Arg Ala Lys Thr Lys
435 440 445Asp Asn Asn Leu Leu Gly Lys Phe Glu Leu Ser Gly Ile Pro
Pro Ala 450 455 460Pro Arg Gly Val Pro Gln Ile Glu Val Thr Phe Asp
Val Asp Ser Asn465 470 475 480Gly Ile Leu Asn Val Ser Ala Val Glu
Lys Gly Thr Gly Lys Ser Asn 485 490 495Lys Ile Thr Ile Thr Asn Asp
Lys Gly Arg Leu Ser Lys Glu Asp Ile 500 505 510Glu Lys Met Val Ala
Glu Ala Glu Lys Phe Lys Glu Glu Asp Glu Lys 515 520 525Glu Ser Gln
Arg Ile Ala Ser Lys Asn Gln Leu Glu Ser Ile Ala Tyr 530 535 540Ser
Leu Lys Asn Thr Ile Ser Glu Ala Gly Asp Lys Leu Glu Gln Ala545 550
555 560Asp Lys Asp Ala Val Thr Lys Lys Ala Glu Glu Thr Ile Ala Trp
Leu 565 570 575Asp Ser Asn Thr Thr Ala Thr Lys Glu Glu Phe Asp Asp
Gln Leu Lys 580 585 590Glu Leu Gln Glu Val Ala Asn Pro Ile Met Ser
Lys Leu Tyr Gln Ala 595 600 605Gly Gly Ala Pro Glu Gly Ala Ala Pro
Gly Gly Phe Pro Gly Gly Ala 610 615 620Pro Pro Ala Pro Glu Ala Glu
Gly Pro Thr Val Glu Glu Val Asp625 630 635161920DNASaccharomyces
cerevisiae 16atgtctaaag ctgtcggtat tgatttaggt actacctact cctgtgttgc
tcacttctct 60aatgatcgtg ttgacattat tgccaacgac caaggtaaca gaaccactcc
atctttcgtt 120ggtttcactg atactgaaag attgattggt gacgctgcta
agaaccaagc tgctatgaac 180ccagctaaca ctgttttcga cgctaagcgt
ttgatcggta gaaacttcaa tgacccagaa 240gtccaaggtg atatgaagca
cttcccattc aagttgatcg atgttgacgg taagccacaa 300attcaagttg
aatttaaggg tgaaaccaag aactttaccc cagaacaaat ctcctccatg
360gtcttgggta agatgaagga aactgccgaa tcttacttgg gtgccaaggt
caatgacgct 420gtcgtcactg tcccagctta cttcaacgat tctcaaagac
aagctaccaa ggatgctggt 480accattgctg gtttgaatgt cttgcgtatt
attaacgaac ctaccgccgc tgccattgct 540tacggtttgg acaagaaggg
taaggaagaa cacgtcttga ttttcgactt gggtggtggt 600actttcgatg
tctctttgtt gtccattgaa gacggtatct ttgaagttaa ggccaccgct
660ggtgacaccc atttgggtgg tgaagatttt gacaacagat tggtcaacca
cttcatccaa 720gaattcaaga gaaagaacaa gaaggacttg tctaccaacc
aaagagcttt gagaagatta 780agaactgctt gtgaaagagc caagagaact
ttgtcttcct ccgctcaaac ttccgttgaa 840attgactctt tgttcgaagg
tatcgatttc tacacttcca tcaccagagc cagattcgaa 900gaattgtgtg
ctgacttgtt cagatctact ttggacccag ttgaaaaggt cttgagagat
960gctaaattgg ataaatctca agtcgatgaa attgtcttgg tcggtggttc
taccagaatt 1020ccaaaggtcc aaaaattggt cactgactac ttcaacggta
aggaaccaaa cagatctatc 1080aacccagatg aagctgttgc ttacggtgct
gctgttcaag ctgctatttt gactggtgac 1140gaatcttcca agactcaaga
tctattgttg ttggatgtcg ctccattatc cttgggtatt 1200gaaactgctg
gtggtgtcat gaccaagttg attccaagaa actctaccat tccaactaag
1260aaatccgaag ttttctctac ttatgctgac aaccaaccag gtgtcttgat
tcaagtcttt 1320gaaggtgaaa gagccaagac taaggacaac aacttgttgg
gtaagttcga attgagtggt 1380attccaccag ctccaagagg tgtcccacaa
attgaagtca ctttcgatgt cgactctaac 1440ggtattttga atgtttccgc
cgtcgaaaag ggtactggta agtctaacaa gatcactatt 1500accaacgaca
agggtagatt gtccaaggaa gatatcgaaa agatggttgc tgaagccgaa
1560aaattcaagg aagaagatga aaaggaatct caaagaattg cttccaagaa
ccaattggaa 1620tccattgctt actctttgaa gaacaccatt tctgaagctg
gtgacaagct agagcaagct 1680gacaaggacg ctgtcactaa gaaggctgaa
gaaactattg cttggttaga cagcaacacc 1740actgctacca aggaagaatt
cgatgaccaa ttgaaggaat tgcaagaggt tgccaaccca 1800atcatgtcta
aattgtacca agctggtggt gctccagaag gcgcagctcc aggtggtttc
1860ccaggtggtg ctcctccagc tccagaagct gaaggtccaa ctgtcgaaga
agttgattaa 192017649PRTSaccharomyces cerevisiae 17Met Ser Arg Ala
Val Gly Ile Asp Leu Gly Thr Thr Tyr Ser Cys Val1 5 10 15Ala His Phe
Ser Asn Asp Arg Val Glu Ile Ile Ala Asn Asp Gln Gly 20 25 30Asn Arg
Thr Thr Pro Ser Tyr Val Ala Phe Thr Asp Thr Glu Arg Leu 35 40 45Ile
Gly Asp Ala Ala Lys Asn Gln Ala Ala Ile Asn Pro His Asn Thr 50 55
60Val Phe Asp Ala Lys Arg Leu Ile Gly Arg Lys Phe Asp Asp Pro Glu65
70 75 80Val Thr Thr Asp Ala Lys His Phe Pro Phe Lys Val Ile Ser Arg
Asp 85 90 95Gly Lys Pro Val Val Gln Val Glu Tyr Lys Gly Glu Thr Lys
Thr Phe 100 105 110Thr Pro Glu Glu Ile Ser Ser Met Val Leu Ser Lys
Met Lys Glu Thr 115 120 125Ala Glu Asn Tyr Leu Gly Thr Thr Val Asn
Asp Ala Val Val Thr Val 130 135 140Pro Ala Tyr Phe Asn Asp Ser Gln
Arg Gln Ala Thr Lys Asp Ala Gly145 150 155 160Thr Ile Ala Gly Met
Asn Val Leu Arg Ile Ile Asn Glu Pro Thr Ala 165 170 175Ala Ala Ile
Ala Tyr Gly Leu Asp Lys Lys Gly Arg Ala Glu His Asn 180 185 190Val
Leu Ile Phe Asp Leu Gly Gly Gly Thr Phe Asp Val Ser Leu Leu 195 200
205Ser Ile Asp Glu Gly Val Phe Glu Val Lys Ala Thr Ala Gly Asp Thr
210 215 220His Leu Gly Gly Glu Asp Phe Asp Asn Arg Leu Val Asn His
Leu Ala225 230 235 240Thr Glu Phe Lys Arg Lys Thr Lys Lys Asp Ile
Ser Asn Asn Gln Arg 245 250 255Ser Leu Arg Arg Leu Arg Thr Ala Ala
Glu Arg Ala Lys Arg Ala Leu 260 265 270Ser Ser Ser Ser Gln Thr Ser
Ile Glu Ile Asp Ser Leu Phe Glu Gly 275 280 285Met Asp Phe Tyr Thr
Ser Leu Thr Arg Ala Arg Phe Glu Glu Leu Cys 290 295 300Ala Asp Leu
Phe Arg Ser Thr Leu Glu Pro Val Glu Lys Val Leu Lys305 310 315
320Asp Ser Lys Leu Asp Lys Ser Gln Ile Asp Glu Ile Val Leu Val Gly
325 330 335Gly Ser Thr Arg Ile Pro Lys Ile Gln Lys Leu Val Ser Asp
Phe Phe 340 345 350Asn Gly Lys Glu Pro Asn Arg Ser Ile Asn Pro Asp
Glu Ala Val Ala 355 360 365Tyr Gly Ala Ala Val Gln Ala Ala Ile Leu
Thr Gly Asp Gln Ser Thr 370 375 380Lys Thr Gln Asp Leu Leu Leu Leu
Asp Val Ala Pro Leu Ser Leu Gly385 390 395 400Ile Glu Thr Ala Gly
Gly Ile Met Thr Lys Leu Ile Pro Arg Asn Ser 405 410 415Thr Ile Pro
Thr Lys Lys Ser Glu Thr Phe Ser Thr Tyr Ala Asp Asn 420 425 430Gln
Pro Gly Val Leu Ile Gln Val Phe Glu Gly Glu Arg Thr Arg Thr 435 440
445Lys Asp Asn Asn Leu Leu Gly Lys Phe Glu Leu Ser Gly Ile Pro Pro
450 455 460Ala Pro Arg Gly Val Pro Gln Ile Asp Val Thr Phe Asp Ile
Asp Ala465 470 475 480Asn Gly Ile Leu Asn Val Ser Ala Leu Glu Lys
Gly Thr Gly Lys Ser 485 490 495Asn Lys Ile Thr Ile Thr Asn Asp Lys
Gly Arg Leu Ser Lys Asp Asp 500 505 510Ile Asp Arg Met Val Ser Glu
Ala Glu Lys Tyr Arg Ala Asp Asp Glu 515 520 525Arg Glu Ala Glu Arg
Val Gln Ala Lys Asn Gln Leu Glu Ser Tyr Ala 530 535 540Phe Thr Leu
Lys Asn Thr Ile Asn Glu Ala Ser Phe Lys Glu Lys Val545 550 555
560Gly Glu Asp Asp Ala Lys Arg Leu Glu Thr Ala Ser Gln Glu Thr Ile
565 570 575Asp Trp Leu Asp Ala Ser Gln Ala Ala Ser Thr Asp Glu Tyr
Lys Asp 580 585 590Arg Gln Lys Glu Leu Glu Gly Ile Ala Asn Pro Ile
Met Thr Lys Phe 595 600 605Tyr Gly Ala Gly Ala Gly Ala Gly Pro Gly
Ala Gly Glu Ser Gly Gly 610 615 620Phe Pro Gly Ser Met Pro Asn Ser
Gly Ala Thr Gly Gly Gly Glu Asp625 630 635 640Thr Gly Pro Thr Val
Glu Glu Val Asp 645181950DNASaccharomyces cerevisiae 18atgtctagag
cagttggtat tgatttggga acaacttact cgtgtgttgc tcatttttcc 60aatgataggg
tagagataat tgcaaatgat caaggtaata ggaccactcc atcgtatgtg
120gctttcacag acaccgaaag attaattggt gacgccgcca aaaatcaagc
tgcaatcaat 180cctcataata cagtttttga tgcaaagcgg ttaattggtc
gtaaatttga
tgatcctgaa 240gtgacgacag atgccaagca cttccctttc aaagttatat
ccagagatgg taaacctgta 300gtgcaagtag aatataaggg tgaaacgaaa
acatttacgc ctgaggaaat ttcttccatg 360gttttaagca aaatgaagga
aactgctgag aactatttgg gaactacggt caatgatgct 420gttgtaactg
ttcctgcata tttcaatgat tctcaaagac aagccactaa ggatgcagga
480actattgcag ggatgaacgt tttacgtatt atcaatgaac ccactgcagc
agcaattgct 540tatggcttgg ataagaaagg cagggctgag cacaatgtcc
tgatttttga tttgggtggt 600ggtacttttg acgtctcttt actttcaatt
gatgagggtg tttttgaggt taaggctacc 660gcaggagaca ctcatttagg
tggtgaagat tttgataata ggttggtgaa ccatttagcc 720actgaattca
aaaggaaaac gaaaaaggac atctctaata atcaaagatc gttaagaaga
780ttgagaactg cggcagaaag agctaagaga gcgctttctt cctcatctca
aacctcgatc 840gagatcgatt ctttatttga aggtatggat ttctacactt
cgttaacaag ggcaaggttt 900gaagagctat gtgctgattt attcagatcc
acattggaac cagtagaaaa ggttcttaaa 960gattcgaagc tggacaagtc
ccaaattgat gagattgtgt tagtcggtgg atctaccaga 1020atcccaaaga
ttcagaaatt agtttctgac ttcttcaatg gcaaagagcc taatcgttct
1080atcaacccgg atgaggctgt tgcttatggt gcagccgttc aagctgccat
tttaaccggc 1140gatcaatcaa caaagacaca agatttacta ttattggatg
ttgcgccatt gtccctagga 1200attgaaactg caggcggcat aatgactaag
ctaattccta gaaactcaac gattccaaca 1260aagaaatcgg aaaccttctc
tacctatgca gataatcaac ctggtgtttt aattcaagtc 1320tttgaaggtg
aaagaacaag aacaaaggat aataacttac ttggtaaatt cgaattaagt
1380ggcattccgc ctgctcccag aggtgtgcct caaattgatg ttacctttga
tatcgacgct 1440aatggtattc ttaatgtgtc tgctttggaa aagggtactg
gtaagagtaa caaaatcacg 1500atcactaacg ataaaggtag gctctcgaag
gatgatattg ataggatggt ttctgaagct 1560gaaaaatata gggctgacga
tgaaagggag gcagaacgag ttcaggctaa gaatcagctt 1620gaatcgtatg
catttacttt gaagaatacc ataaacgaag caagtttcaa agagaaagta
1680ggtgaagatg atgcaaagag attagaaaca gcgtctcagg aaaccattga
ctggttagat 1740gcatcgcagg cagcctctac ggacgaatat aaggatagac
aaaaggagtt ggaaggcatt 1800gccaatccaa taatgacgaa attttacggt
gctggtgccg gcgcaggtcc tggagcgggg 1860gaatccggtg gattccccgg
atccatgccc aactcgggtg ctacgggagg tggagaagat 1920acaggtccaa
cagtggaaga ggttgattga 195019642PRTSaccharomyces cerevisiae 19Met
Ser Lys Ala Val Gly Ile Asp Leu Gly Thr Thr Tyr Ser Cys Val1 5 10
15Ala His Phe Ala Asn Asp Arg Val Glu Ile Ile Ala Asn Asp Gln Gly
20 25 30Asn Arg Thr Thr Pro Ser Tyr Val Ala Phe Thr Asp Thr Glu Arg
Leu 35 40 45Ile Gly Asp Ala Ala Lys Asn Gln Ala Ala Met Asn Pro His
Asn Thr 50 55 60Val Phe Asp Ala Lys Arg Leu Ile Gly Arg Lys Phe Asp
Asp Pro Glu65 70 75 80Val Thr Asn Asp Ala Lys His Tyr Pro Phe Lys
Val Ile Asp Lys Gly 85 90 95Gly Lys Pro Val Val Gln Val Glu Tyr Lys
Gly Glu Thr Lys Thr Phe 100 105 110Thr Pro Glu Glu Ile Ser Ser Met
Ile Leu Thr Lys Met Lys Glu Thr 115 120 125Ala Glu Asn Phe Leu Gly
Thr Glu Val Lys Asp Ala Val Val Thr Val 130 135 140Pro Ala Tyr Phe
Asn Asp Ser Gln Arg Gln Ala Thr Lys Asp Ala Gly145 150 155 160Thr
Ile Ala Gly Leu Asn Val Leu Arg Ile Ile Asn Glu Pro Thr Ala 165 170
175Ala Ala Ile Ala Tyr Gly Leu Asp Lys Lys Ser Gln Lys Glu His Asn
180 185 190Val Leu Ile Phe Asp Leu Gly Gly Gly Thr Phe Asp Val Ser
Leu Leu 195 200 205Ser Ile Asp Glu Gly Val Phe Glu Val Lys Ala Thr
Ala Gly Asp Thr 210 215 220His Leu Gly Gly Glu Asp Phe Asp Ser Arg
Leu Val Asn Phe Leu Ala225 230 235 240Glu Glu Phe Lys Arg Lys Asn
Lys Lys Asp Leu Thr Thr Asn Gln Arg 245 250 255Ser Leu Arg Arg Leu
Arg Thr Ala Ala Glu Arg Ala Lys Arg Thr Leu 260 265 270Ser Ser Ser
Ala Gln Thr Ser Ile Glu Ile Asp Ser Leu Phe Glu Gly 275 280 285Ile
Asp Phe Tyr Thr Ser Ile Thr Arg Ala Arg Phe Glu Glu Leu Cys 290 295
300Ala Asp Leu Phe Arg Ser Thr Leu Glu Pro Val Glu Lys Val Leu
Ala305 310 315 320Asp Ser Lys Leu Asp Lys Ser Gln Ile Asp Glu Ile
Val Leu Val Gly 325 330 335Gly Ser Thr Arg Ile Pro Lys Val Gln Lys
Leu Val Ser Asp Phe Phe 340 345 350Asn Gly Lys Glu Pro Asn Arg Ser
Ile Asn Pro Asp Glu Ala Val Ala 355 360 365Tyr Gly Ala Ala Val Gln
Ala Ala Ile Leu Thr Gly Asp Gln Ser Ser 370 375 380Thr Thr Gln Asp
Leu Leu Leu Leu Asp Val Ala Pro Leu Ser Leu Gly385 390 395 400Ile
Glu Thr Ala Gly Gly Ile Met Thr Lys Leu Ile Pro Arg Asn Ser 405 410
415Thr Ile Pro Thr Lys Lys Ser Glu Val Phe Ser Thr Tyr Ala Asp Asn
420 425 430Gln Pro Gly Val Leu Ile Gln Val Phe Glu Gly Glu Arg Thr
Arg Thr 435 440 445Lys Asp Asn Asn Leu Leu Gly Lys Phe Glu Leu Ser
Gly Ile Pro Pro 450 455 460Ala Pro Arg Gly Val Pro Gln Ile Glu Val
Thr Phe Asp Ile Asp Ala465 470 475 480Asn Gly Ile Leu Asn Val Ser
Ala Val Glu Lys Gly Thr Gly Lys Ser 485 490 495Asn Lys Ile Thr Ile
Thr Asn Asp Lys Gly Arg Leu Ser Lys Glu Asp 500 505 510Ile Asp Lys
Met Val Ala Glu Ala Glu Lys Phe Lys Ala Glu Asp Glu 515 520 525Gln
Glu Ala Gln Arg Val Gln Ala Lys Asn Gln Leu Glu Ser Tyr Ala 530 535
540Phe Thr Leu Lys Asn Ser Val Ser Glu Asn Asn Phe Lys Glu Lys
Val545 550 555 560Gly Glu Glu Asp Ala Arg Lys Leu Glu Ala Ala Ala
Gln Asp Ala Ile 565 570 575Asn Trp Leu Asp Ala Ser Gln Ala Ala Ser
Thr Glu Glu Tyr Lys Glu 580 585 590Arg Gln Lys Glu Leu Glu Gly Val
Ala Asn Pro Ile Met Ser Lys Phe 595 600 605Tyr Gly Ala Ala Gly Gly
Ala Pro Gly Ala Gly Pro Val Pro Gly Ala 610 615 620Gly Ala Gly Pro
Thr Gly Ala Pro Asp Asn Gly Pro Thr Val Glu Glu625 630 635 640Val
Asp201929DNASaccharomyces cerevisiae 20atgtcaaaag ctgttggtat
tgatttaggt acaacctatt catgtgttgc tcattttgca 60aacgataggg ttgaaattat
cgctaacgat caaggtaata gaacgacgcc ttcttatgtg 120gcttttactg
acacagaaag gctaattggt gacgctgcga agaatcaagc tgcgatgaac
180ccacataata cagtattcga tgctaagcgt ctgatcggac gtaaattcga
tgatccagaa 240gtgacgaacg atgctaagca ttacccattc aaagtgattg
acaagggagg taaaccggta 300gtgcaagtgg aatataaagg cgagacaaag
acatttactc cagaagaaat ttcctcaatg 360atcttgacaa agatgaagga
gactgctgag aactttttag gaacagaagt gaaagatgct 420gtagtaacgg
ttccagccta tttcaacgat tcacaaaggc aagcaacaaa agatgccggt
480acaatcgcgg gcttgaacgt tcttcgtatc attaatgaac ctacagctgc
cgctattgcg 540tatgggctgg acaagaaatc gcagaaggag cacaacgtct
tgatctttga tttaggtggt 600ggtacttttg atgtctctct gctatccata
gatgaaggtg tctttgaggt taaggctact 660gctggtgaca ctcacttggg
tggtgaagat ttcgatagta ggctggttaa ctttctagcc 720gaggagttca
aaagaaaaaa taaaaaggat ctaacaacta accaaaggtc cctaaggagg
780ttaaggaccg ccgctgaaag ggccaagaga actctgtctt cgtctgctca
gacatctata 840gaaatagatt cattatttga gggtatcgat ttctatactt
ccattacaag ggcaagattt 900gaagaattat gtgctgattt gtttagatct
acattggagc cagtggaaaa agttttggct 960gattcaaaat tagataagtc
acaaattgat gaaattgtac ttgttggtgg ttcaacaaga 1020attccaaaag
tacaaaaact ggtttctgat tttttcaatg gtaaagaacc aaaccgttcg
1080attaaccctg atgaggccgt cgcttatggt gctgccgtac aggctgccat
cttaacgggt 1140gaccagtcgt cgacgaccca agatttactg ttgctggatg
ttgcaccatt atctctaggt 1200attgaaactg caggtggtat tatgacaaag
ttgatcccaa gaaattcgac tatcccaaca 1260aaaaaatcgg aagtgttttc
cacctacgct gacaaccaac ctggtgtgtt gatacaagtt 1320tttgagggtg
aaaggacaag gacaaaagac aacaatctac tgggtaaatt tgagttgagc
1380ggtattccac ccgctccaag aggcgtacca caaattgaag ttacatttga
tatcgatgca 1440aatggtattc tgaacgtatc tgccgttgaa aaaggtactg
gtaaatctaa caagattaca 1500attactaacg ataagggaag attatcgaag
gaagatatcg ataaaatggt tgctgaggca 1560gaaaagttca aggccgaaga
tgaacaagaa gctcaacgtg ttcaagctaa gaatcagcta 1620gaatcgtacg
cgtttacttt gaaaaattct gtgagcgaaa ataacttcaa ggagaaggtg
1680ggtgaagagg atgccaggaa attggaagcc gccgcccaag atgctataaa
ttggttagat 1740gcttcgcaag cggcctccac cgaggaatac aaggaaaggc
aaaaggaact agaaggtgtt 1800gcaaacccca ttatgagtaa attttacgga
gctgcaggtg gtgccccagg agcaggccca 1860gttccgggtg ctggagcagg
ccccactgga gcaccagaca acggcccaac ggttgaagag 1920gttgattag
192921693PRTSaccharomyces cerevisiae 21Met Ser Thr Pro Phe Gly Leu
Asp Leu Gly Asn Asn Asn Ser Val Leu1 5 10 15Ala Val Ala Arg Asn Arg
Gly Ile Asp Ile Val Val Asn Glu Val Ser 20 25 30Asn Arg Ser Thr Pro
Ser Val Val Gly Phe Gly Pro Lys Asn Arg Tyr 35 40 45Leu Gly Glu Thr
Gly Lys Asn Lys Gln Thr Ser Asn Ile Lys Asn Thr 50 55 60Val Ala Asn
Leu Lys Arg Ile Ile Gly Leu Asp Tyr His His Pro Asp65 70 75 80Phe
Glu Gln Glu Ser Lys His Phe Thr Ser Lys Leu Val Glu Leu Asp 85 90
95Asp Lys Lys Thr Gly Ala Glu Val Arg Phe Ala Gly Glu Lys His Val
100 105 110Phe Ser Ala Thr Gln Leu Ala Ala Met Phe Ile Asp Lys Val
Lys Asp 115 120 125Thr Val Lys Gln Asp Thr Lys Ala Asn Ile Thr Asp
Val Cys Ile Ala 130 135 140Val Pro Pro Trp Tyr Thr Glu Glu Gln Arg
Tyr Asn Ile Ala Asp Ala145 150 155 160Ala Arg Ile Ala Gly Leu Asn
Pro Val Arg Ile Val Asn Asp Val Thr 165 170 175Ala Ala Gly Val Ser
Tyr Gly Ile Phe Lys Thr Asp Leu Pro Glu Gly 180 185 190Glu Glu Lys
Pro Arg Ile Val Ala Phe Val Asp Ile Gly His Ser Ser 195 200 205Tyr
Thr Cys Ser Ile Met Ala Phe Lys Lys Gly Gln Leu Lys Val Leu 210 215
220Gly Thr Ala Cys Asp Lys His Phe Gly Gly Arg Asp Phe Asp Leu
Ala225 230 235 240Ile Thr Glu His Phe Ala Asp Glu Phe Lys Thr Lys
Tyr Lys Ile Asp 245 250 255Ile Arg Glu Asn Pro Lys Ala Tyr Asn Arg
Ile Leu Thr Ala Ala Glu 260 265 270Lys Leu Lys Lys Val Leu Ser Ala
Asn Thr Asn Ala Pro Phe Ser Val 275 280 285Glu Ser Val Met Asn Asp
Val Asp Val Ser Ser Gln Leu Ser Arg Glu 290 295 300Glu Leu Glu Glu
Leu Val Lys Pro Leu Leu Glu Arg Val Thr Glu Pro305 310 315 320Val
Thr Lys Ala Leu Ala Gln Ala Lys Leu Ser Ala Glu Glu Val Asp 325 330
335Phe Val Glu Ile Ile Gly Gly Thr Thr Arg Ile Pro Thr Leu Lys Gln
340 345 350Ser Ile Ser Glu Ala Phe Gly Lys Pro Leu Ser Thr Thr Leu
Asn Gln 355 360 365Asp Glu Ala Ile Ala Lys Gly Ala Ala Phe Ile Cys
Ala Ile His Ser 370 375 380Pro Thr Leu Arg Val Arg Pro Phe Lys Phe
Glu Asp Ile His Pro Tyr385 390 395 400Ser Val Ser Tyr Ser Trp Asp
Lys Gln Val Glu Asp Glu Asp His Met 405 410 415Glu Val Phe Pro Ala
Gly Ser Ser Phe Pro Ser Thr Lys Leu Ile Thr 420 425 430Leu Asn Arg
Thr Gly Asp Phe Ser Met Ala Ala Ser Tyr Thr Asp Ile 435 440 445Thr
Gln Leu Pro Pro Asn Thr Pro Glu Gln Ile Ala Asn Trp Glu Ile 450 455
460Thr Gly Val Gln Leu Pro Glu Gly Gln Asp Ser Val Pro Val Lys
Leu465 470 475 480Lys Leu Arg Cys Asp Pro Ser Gly Leu His Thr Ile
Glu Glu Ala Tyr 485 490 495Thr Ile Glu Asp Ile Glu Val Glu Glu Pro
Ile Pro Leu Pro Glu Asp 500 505 510Ala Pro Glu Asp Ala Glu Gln Glu
Phe Lys Lys Val Thr Lys Thr Val 515 520 525Lys Lys Asp Asp Leu Thr
Ile Val Ala His Thr Phe Gly Leu Asp Ala 530 535 540Lys Lys Leu Asn
Glu Leu Ile Glu Lys Glu Asn Glu Met Leu Ala Gln545 550 555 560Asp
Lys Leu Val Ala Glu Thr Glu Asp Arg Lys Asn Thr Leu Glu Glu 565 570
575Tyr Ile Tyr Thr Leu Arg Gly Lys Leu Glu Glu Glu Tyr Ala Pro Phe
580 585 590Ala Ser Asp Ala Glu Lys Thr Lys Leu Gln Gly Met Leu Asn
Lys Ala 595 600 605Glu Glu Trp Leu Tyr Asp Glu Gly Phe Asp Ser Ile
Lys Ala Lys Tyr 610 615 620Ile Ala Lys Tyr Glu Glu Leu Ala Ser Leu
Gly Asn Ile Ile Arg Gly625 630 635 640Arg Tyr Leu Ala Lys Glu Glu
Glu Lys Lys Gln Ala Ile Arg Ser Lys 645 650 655Gln Glu Ala Ser Gln
Met Ala Ala Met Ala Glu Lys Leu Ala Ala Gln 660 665 670Arg Lys Ala
Glu Ala Glu Lys Lys Glu Glu Lys Lys Asp Thr Glu Gly 675 680 685Asp
Val Asp Met Asp 690222082DNASaccharomyces cerevisiae 22atgagtactc
catttggttt agatttaggt aacaataact ctgtccttgc cgttgctaga 60aacagaggta
tcgacattgt cgttaatgaa gtctctaacc gttccacccc atctgttgtt
120ggttttggtc caaagaacag atacttgggt gaaactggta agaacaagca
gacttccaac 180atcaagaaca ctgtcgccaa cttgaaaaga attattggtt
tggattacca ccatccagat 240ttcgagcaag aatctaagca cttcacctct
aagttggttg aattggatga caagaagact 300ggtgccgaag ttagattcgc
tggtgagaaa catgtttttt cagctactca actagctgcc 360atgttcatcg
acaaagtcaa ggacaccgtc aagcaggaca caaaggcaaa tattaccgat
420gtttgtattg ctgtcccacc ttggtacacc gaagaacaac gttacaacat
tgctgatgct 480gctagaattg ctggtttgaa ccctgttaga attgtcaacg
acgttactgc tgccggtgtt 540tcttacggta tcttcaagac tgatttgcct
gaaggcgaag aaaagccaag aattgttgcc 600tttgttgata ttggtcactc
ttcctacacc tgttctatca tggccttcaa gaagggtcaa 660ttgaaagtct
taggaactgc ctgcgacaag cattttggtg gtagggactt cgatttggct
720ataacagaac atttcgccga tgagttcaaa actaaataca agattgacat
cagagaaaat 780ccaaaggctt acaacagaat tctaactgct gctgaaaagt
tgaagaaagt tttgtctgct 840aatactaatg ccccattctc tgttgaatcc
gtcatgaacg acgttgatgt ttcctctcaa 900ttatctcgtg aagaattaga
agaattggtc aagccattgt tggaacgtgt tactgaacca 960gttaccaaag
ctttagctca agccaaatta tctgctgaag aagttgattt tgttgaaatt
1020attggtggta ctactcgtat cccaacattg aaacaatcca tttctgaagc
cttcggcaag 1080ccattgtcca ccactttgaa ccaagatgaa gccatcgcca
agggtgccgc ctttatttgc 1140gccattcact ctccaactct aagagttaga
ccattcaagt ttgaggatat ccatccttac 1200tctgtctctt actcttggga
caagcaagtt gaggacgaag accacatgga agttttccca 1260gctggttcat
ccttcccatc tactaaattg atcactttga accgtacggg tgacttttca
1320atggctgcta gctacactga catcacacag ttaccaccaa acactccaga
acaaatcgct 1380aactgggaga tcactggtgt tcaattacca gaaggtcaag
actctgttcc tgttaagtta 1440aagttgagat gcgacccctc tggtttacac
acaattgaag aggcttacac tattgaagat 1500attgaagttg aagaacctat
tccattacca gaagatgctc cagaagatgc tgagcaagaa 1560tttaagaagg
ttactaaaac tgtaaagaag gatgacttaa ccatcgttgc acacaccttt
1620ggcctagacg ctaaaaagtt gaatgaatta attgaaaaag aaaatgaaat
gcttgctcaa 1680gataagctag ttgctgagac agaagaccgt aagaacactc
ttgaagagta catctacaca 1740ttgcgtggta agttggaaga agagtatgct
ccatttgctt ccgatgctga aaagacgaag 1800ttacaaggta tgttaaacaa
ggccgaagag tggttatacg atgaaggttt cgattccatc 1860aaagctaagt
acattgccaa atacgaagaa ttggcttctc taggtaacat tattagaggt
1920agatacttgg ctaaagaaga agaaaagaag caagctataa gatctaagca
agaagcatcc 1980caaatggctg ctatggctga aaagttggct gctcaaagaa
aggcagaagc tgaaaagaag 2040gaagaaaaga aggacactga aggtgatgtt
gacatggact aa 208223693PRTSaccharomyces cerevisiae 23Met Ser Thr
Pro Phe Gly Leu Asp Leu Gly Asn Asn Asn Ser Val Leu1 5 10 15Ala Val
Ala Arg Asn Arg Gly Ile Asp Val Val Val Asn Glu Val Ser 20 25 30Asn
Arg Ser Thr Pro Ser Leu Val Gly Phe Gly Pro Arg Asn Arg Tyr 35 40
45Leu Gly Glu Ser Gly Lys Thr Lys Gln Thr Ser Asn Val Lys Asn Thr
50 55 60Val Glu Asn Leu Lys Arg Ile Ile Gly Leu Lys Phe Lys Asp Pro
Glu65 70 75 80Phe Asp Ile Glu Asn Lys Phe Phe Thr Ser Lys Leu Val
Gln Leu Lys 85 90 95Asn Gly Lys Val Gly Val Glu Val Glu Phe Gly Gly
Lys Thr His Val 100 105 110Phe Ser Ala Thr Gln Leu Thr Ala Met Phe
Ile Asp Lys Val Lys His 115 120 125Thr Val Gln Glu Glu Thr Lys Ser
Ser Ile Thr Asp Val Cys Leu Ala 130 135 140Val Pro Val Trp Tyr Ser
Glu Glu Gln Arg Tyr Asn Ile Ala Asp Ala145 150 155
160Ala Arg Ile Ala Gly Leu Asn Pro Val Arg Ile Val Asn Asp Val Thr
165 170 175Ala Ala Ala Val Ser Tyr Gly Val Phe Lys Asn Asp Leu Pro
Gly Pro 180 185 190Glu Glu Lys Pro Arg Ile Ile Gly Leu Val Asp Ile
Gly His Ser Thr 195 200 205Tyr Thr Cys Ser Ile Met Ala Phe Arg Lys
Gly Glu Met Lys Val Leu 210 215 220Gly Thr Ala Tyr Asp Lys His Phe
Gly Gly Arg Asp Phe Asp Arg Ala225 230 235 240Ile Thr Glu His Phe
Ala Asp Gln Phe Lys Asp Lys Tyr Lys Ile Asp 245 250 255Ile Arg Lys
Asn Pro Lys Ala Tyr Asn Arg Ile Leu Ile Ala Ala Glu 260 265 270Lys
Leu Lys Lys Val Leu Ser Ala Asn Thr Thr Ala Pro Phe Ser Val 275 280
285Glu Ser Val Met Asp Asp Ile Asp Val Ser Ser Gln Leu Ser Arg Glu
290 295 300Glu Leu Glu Glu Leu Val Glu Pro Leu Leu Lys Arg Val Thr
Tyr Pro305 310 315 320Ile Thr Asn Ala Leu Ala Gln Ala Lys Leu Thr
Val Asn Asp Ile Asp 325 330 335Phe Val Glu Ile Ile Gly Gly Thr Thr
Arg Ile Pro Val Leu Lys Lys 340 345 350Ser Ile Ser Asp Val Phe Gly
Lys Pro Leu Ser Ser Thr Leu Asn Gln 355 360 365Asp Glu Ala Val Ala
Lys Gly Ala Ala Phe Ile Cys Ala Ile His Ser 370 375 380Pro Thr Leu
Arg Val Arg Pro Phe Lys Phe Glu Asp Ile Asp Pro Tyr385 390 395
400Ser Val Ser Tyr Thr Trp Asp Lys Gln Val Asp Asp Glu Asp Arg Leu
405 410 415Glu Val Phe Pro Ala Asn Ser Ser Tyr Pro Ser Thr Lys Leu
Ile Thr 420 425 430Leu His Arg Thr Gly Asp Phe Ser Met Lys Ala Val
Tyr Thr His Pro 435 440 445Ser Lys Leu Pro Lys Gly Thr Ser Thr Thr
Ile Ala Lys Trp Ser Phe 450 455 460Thr Gly Val Lys Val Pro Lys Asp
Gln Asp Phe Ile Pro Val Lys Val465 470 475 480Lys Leu Arg Cys Asp
Pro Ser Gly Leu His Ile Ile Glu Asn Ala Tyr 485 490 495Thr Thr Glu
Asp Ile Thr Val Gln Glu Pro Val Pro Leu Pro Glu Asp 500 505 510Ala
Pro Glu Asp Ala Glu Pro Gln Phe Lys Glu Val Thr Lys Thr Ile 515 520
525Lys Lys Asp Val Leu Gly Met Thr Ala Lys Thr Phe Ala Leu Asn Pro
530 535 540Val Glu Leu Asn Asp Leu Ile Glu Lys Glu Asn Glu Leu Arg
Asn Gln545 550 555 560Asp Lys Leu Val Ala Glu Thr Glu Asp Arg Lys
Asn Ala Leu Glu Glu 565 570 575Tyr Ile Tyr Thr Leu Arg Ala Lys Leu
Asp Asp Glu Tyr Ser Asp Phe 580 585 590Ala Ser Asp Ala Glu Lys Glu
Lys Leu Lys Asn Met Leu Ala Thr Thr 595 600 605Glu Asn Trp Leu Tyr
Gly Asp Gly Asp Asp Ser Thr Lys Ala Lys Tyr 610 615 620Ile Ala Lys
Tyr Glu Glu Leu Ala Ser Leu Gly Asn Ile Ile Arg Gly625 630 635
640Arg Tyr Leu Ala Lys Glu Glu Glu Lys Arg Gln Ala Leu Arg Ala Asn
645 650 655Gln Glu Thr Ser Lys Met Asn Asp Ile Ala Glu Lys Leu Ala
Glu Gln 660 665 670Arg Arg Ala Arg Ala Ala Ser Asp Asp Ser Asp Asp
Asn Asn Asp Glu 675 680 685Asn Met Asp Leu Asp
690242082DNASaccharomyces cerevisiae 24atgagcactc catttggctt
agatttaggt aacaataact cagtactagc agttgccaga 60aataggggta ttgatgtcgt
tgtcaatgaa gtttctaata ggtctacacc atccttggtc 120ggctttggcc
ccagaaatag gtacttaggt gaatctggta aaactaagca aacatcgaat
180gttaaaaaca ctgtggaaaa cttgaaaaga atcattggac taaagttcaa
agaccctgaa 240tttgatatcg agaataagtt cttcacttcg aaattggtac
agctaaaaaa tggtaaagtt 300ggtgtggaag tggagttcgg cggtaaaaca
cacgtatttt cagctactca actgactgct 360atgttcattg ataaggtgaa
gcacaccgtt caagaggaaa cgaagtcatc aattaccgat 420gtctgcctcg
cagttcctgt atggtattcg gaagaacaac gttataacat agccgatgct
480gccagaattg caggattaaa tcctgtaagg attgtcaacg atgtgactgc
agccgccgtt 540tcgtacggcg tcttcaagaa tgatctgcca ggtcctgaag
aaaagccaag aatcattggc 600ttagtggaca ttgggcattc tacctacacc
tgttctatta tggctttccg caaaggcgaa 660atgaaagtat taggtactgc
ttatgacaag cactttggtg gtagagattt cgatcgcgca 720atcacagaac
attttgctga tcagtttaag gacaagtaca agattgacat taggaaaaat
780ccgaaagctt ataacagaat tttaatcgct gctgaaaaat taaaaaaagt
gctttctgcg 840aacactactg cccccttctc cgttgaatct gttatggatg
atatcgacgt ttcctctcaa 900ttgagccgtg aagagctgga agaattagta
gagcccttgt tgaagcgtgt gacgtatcca 960atcaccaatg cattggctca
agctaaatta actgtcaatg atattgactt cgtagaaata 1020attggtggta
caacccgtat cccagtttta aagaagtcaa tttctgatgt ttttggaaaa
1080cctttgtcat ctactttaaa tcaagacgaa gctgtggcca agggggccgc
tttcatatgt 1140gccattcact ctccaacttt aagggtcagg ccgtttaaat
ttgaagatat tgatccgtat 1200tcagtgtcat acacttggga taagcaggtc
gatgacgaag accgtttgga agtattccct 1260gctaattcat catatccatc
aactaaacta attactttac atcgtactgg agatttcagc 1320atgaaagcgg
tgtacactca tccttcgaaa ctgccaaaag gtacttccac cactattgca
1380aaatggagct tcactggggt caaggttcct aaagatcaag attttattcc
tgtaaaggtc 1440aagttaagat gcgatccttc cggcttgcat attatcgaga
acgcttacac aacggaagat 1500attacggttc aagagccagt gcctttaccg
gaagacgcac cagaagatgc cgagccccag 1560tttaaagaag ttactaaaac
aattaagaaa gatgtgctag gtatgactgc aaaaacattc 1620gcgctaaacc
cggttgagtt gaacgatcta attgaaaaag agaatgaatt aagaaaccag
1680gataagttag ttgccgaaac cgaggatcgc aaaaatgccc ttgaagagta
tatttatacc 1740cttcgtgcca aactcgatga tgaatactcc gattttgcgt
ctgacgcaga aaaagaaaag 1800ctaaaaaaca tgttagccac tactgaaaat
tggttatatg gtgatggtga cgattctacc 1860aaggcaaaat acattgctaa
atatgaggag ctggcatcgt tggggaatat tattagaggt 1920agatatttag
caaaggagga agaaaaaaga caagcactca gagcgaatca agaaacttct
1980aaaatgaatg atattgctga aaaattggct gagcaaagaa gggcacgcgc
tgcaagtgat 2040gatagcgatg acaacaatga tgaaaacatg gaccttgatt aa
208225613PRTSaccharomyces cerevisiae 25Met Ala Glu Gly Val Phe Gln
Gly Ala Ile Gly Ile Asp Leu Gly Thr1 5 10 15Thr Tyr Ser Cys Val Ala
Thr Tyr Glu Ser Ser Val Glu Ile Ile Ala 20 25 30Asn Glu Gln Gly Asn
Arg Val Thr Pro Ser Phe Val Ala Phe Thr Pro 35 40 45Glu Glu Arg Leu
Ile Gly Asp Ala Ala Lys Asn Gln Ala Ala Leu Asn 50 55 60Pro Arg Asn
Thr Val Phe Asp Ala Lys Arg Leu Ile Gly Arg Arg Phe65 70 75 80Asp
Asp Glu Ser Val Gln Lys Asp Met Lys Thr Trp Pro Phe Lys Val 85 90
95Ile Asp Val Asp Gly Asn Pro Val Ile Glu Val Gln Tyr Leu Glu Glu
100 105 110Thr Lys Thr Phe Ser Pro Gln Glu Ile Ser Ala Met Val Leu
Thr Lys 115 120 125Met Lys Glu Ile Ala Glu Ala Lys Ile Gly Lys Lys
Val Glu Lys Ala 130 135 140Val Ile Thr Val Pro Ala Tyr Phe Asn Asp
Ala Gln Arg Gln Ala Thr145 150 155 160Lys Asp Ala Gly Ala Ile Ser
Gly Leu Asn Val Leu Arg Ile Ile Asn 165 170 175Glu Pro Thr Ala Ala
Ala Ile Ala Tyr Gly Leu Gly Ala Gly Lys Ser 180 185 190Glu Lys Glu
Arg His Val Leu Ile Phe Asp Leu Gly Gly Gly Thr Phe 195 200 205Asp
Val Ser Leu Leu His Ile Ala Gly Gly Val Tyr Thr Val Lys Ser 210 215
220Thr Ser Gly Asn Thr His Leu Gly Gly Gln Asp Phe Asp Thr Asn
Leu225 230 235 240Leu Glu His Phe Lys Ala Glu Phe Lys Lys Lys Thr
Gly Leu Asp Ile 245 250 255Ser Asp Asp Ala Arg Ala Leu Arg Arg Leu
Arg Thr Ala Ala Glu Arg 260 265 270Ala Lys Arg Thr Leu Ser Ser Val
Thr Gln Thr Thr Val Glu Val Asp 275 280 285Ser Leu Phe Asp Gly Glu
Asp Phe Glu Ser Ser Leu Thr Arg Ala Arg 290 295 300Phe Glu Asp Leu
Asn Ala Ala Leu Phe Lys Ser Thr Leu Glu Pro Val305 310 315 320Glu
Gln Val Leu Lys Asp Ala Lys Ile Ser Lys Ser Gln Ile Asp Glu 325 330
335Val Val Leu Val Gly Gly Ser Thr Arg Ile Pro Lys Val Gln Lys Leu
340 345 350Leu Ser Asp Phe Phe Asp Gly Lys Gln Leu Glu Lys Ser Ile
Asn Pro 355 360 365Asp Glu Ala Val Ala Tyr Gly Ala Ala Val Gln Gly
Ala Ile Leu Thr 370 375 380Gly Gln Ser Thr Ser Asp Glu Thr Lys Asp
Leu Leu Leu Leu Asp Val385 390 395 400Ala Pro Leu Ser Leu Gly Val
Gly Met Gln Gly Asp Met Phe Gly Ile 405 410 415Val Val Pro Arg Asn
Thr Thr Val Pro Thr Ile Lys Arg Arg Thr Phe 420 425 430Thr Thr Cys
Ala Asp Asn Gln Thr Thr Val Gln Phe Pro Val Tyr Gln 435 440 445Gly
Glu Arg Val Asn Cys Lys Glu Asn Thr Leu Leu Gly Glu Phe Asp 450 455
460Leu Lys Asn Ile Pro Met Met Pro Ala Gly Glu Pro Val Leu Glu
Ala465 470 475 480Ile Phe Glu Val Asp Ala Asn Gly Ile Leu Lys Val
Thr Ala Val Glu 485 490 495Lys Ser Thr Gly Lys Ser Ser Asn Ile Thr
Ile Ser Asn Ala Val Gly 500 505 510Arg Leu Ser Ser Glu Glu Ile Glu
Lys Met Val Asn Gln Ala Glu Glu 515 520 525Phe Lys Ala Ala Asp Glu
Ala Phe Ala Lys Lys His Glu Ala Arg Gln 530 535 540Arg Leu Glu Ser
Tyr Val Ala Ser Ile Glu Gln Thr Val Thr Asp Pro545 550 555 560Val
Leu Ser Ser Lys Leu Lys Arg Gly Ser Lys Ser Lys Ile Glu Ala 565 570
575Ala Leu Ser Asp Ala Leu Ala Ala Leu Gln Ile Glu Asp Pro Ser Ala
580 585 590Asp Glu Leu Arg Lys Ala Glu Val Gly Leu Lys Arg Val Val
Thr Lys 595 600 605Ala Met Ser Ser Arg 610261842DNASaccharomyces
cerevisiae 26atggctgaag gtgttttcca aggtgctatc ggtatcgatt taggtacaac
ctactcttgt 60gttgctactt acgaatcctc cgttgaaatt attgccaacg aacaaggtaa
cagagtcacc 120ccatctttcg ttgctttcac tccagaagaa agattgattg
gtgatgctgc caagaaccaa 180gctgctttga acccaagaaa cactgtcttc
gatgctaagc gtttgattgg tagaagattc 240gacgacgaat ctgttcaaaa
ggacatgaag acctggcctt tcaaggttat cgacgtcgat 300ggtaacccag
tcatcgaagt ccaatacttg gaagaaacca agactttctc cccacaagaa
360atttccgcta tggttttgac caagatgaag gaaattgctg aagctaagat
tggtaagaag 420gttgaaaagg ccgtcattac tgtcccagct tactttaacg
acgctcaaag acaagctacc 480aaggatgccg gtgccatttc tggtttgaac
gttttgcgta tcatcaacga acctactgcc 540gctgctattg cttacggtct
aggtgctggt aagtccgaaa aggaaagaca tgttttgatt 600ttcgatttgg
gtggtggtac tttcgatgtt tccttgttgc acattgctgg tggtgtttac
660actgttaaat ctacttccgg taacactcac ttgggtggtc aagatttcga
caccaacttg 720ttggaacact tcaaggctga attcaagaag aagactggtt
tggacatctc cgacgatgcc 780agagctttga gaagattgag aactgctgct
gaaagagcta agagaacctt atcttctgtc 840actcaaacta ccgttgaagt
tgactctttg tttgacggtg aagatttcga atcctctttg 900actagagcta
gatttgaaga cttgaacgcc gcattgttca agtctacttt ggaacctgtt
960gaacaagttt tgaaggatgc taagatctct aagtctcaaa tcgacgaagt
tgtcttggtt 1020ggtggttcca ccagaattcc aaaggtccaa aagttgttgt
ctgacttctt tgacggtaag 1080caattggaaa aatctattaa cccagatgaa
gctgttgctt acggtgctgc tgttcaaggt 1140gctatcttga ccggccaatc
cacatctgac gaaaccaagg acttgttgtt gttagatgtt 1200gctccattat
ctctaggtgt tggtatgcaa ggtgacatgt tcggtatcgt tgttccaaga
1260aacactactg ttccaaccat caagagaaga acctttacta catgtgctga
caaccaaacc 1320accgttcaat tcccagtcta ccaaggtgaa cgtgttaact
gtaaagaaaa cactttgttg 1380ggtgaattcg acttgaagaa catcccaatg
atgccagctg gtgaaccagt cttggaagct 1440atcttcgaag ttgatgctaa
cggtatcttg aaggttactg ccgtcgaaaa gtctaccggt 1500aagtcttcta
acatcactat ctctaacgct gttggtagat tgtcttctga agaaattgaa
1560aagatggtta accaagctga agagttcaag gctgccgatg aagcttttgc
caagaagcac 1620gaagctagac aaagattgga atcctacgtt gcctccatcg
aacaaactgt cactgaccca 1680gtcttgtctt ctaaattgaa gagaggttcc
aagtccaaga ttgaagctgc tttgtccgat 1740gctttggctg ctttgcaaat
cgaagaccca tctgctgatg aattgagaaa ggctgaagtt 1800ggtttgaaga
gagttgtcac caaggccatg tcttctcgtt aa 184227613PRTSaccharomyces
cerevisiae 27Met Ala Glu Gly Val Phe Gln Gly Ala Ile Gly Ile Asp
Leu Gly Thr1 5 10 15Thr Tyr Ser Cys Val Ala Thr Tyr Glu Ser Ser Val
Glu Ile Ile Ala 20 25 30Asn Glu Gln Gly Asn Arg Val Thr Pro Ser Phe
Val Ala Phe Thr Pro 35 40 45Gln Glu Arg Leu Ile Gly Asp Ala Ala Lys
Asn Gln Ala Ala Leu Asn 50 55 60Pro Arg Asn Thr Val Phe Asp Ala Lys
Arg Leu Ile Gly Arg Arg Phe65 70 75 80Asp Asp Glu Ser Val Gln Lys
Asp Met Lys Thr Trp Pro Phe Lys Val 85 90 95Ile Asp Val Asp Gly Asn
Pro Val Ile Glu Val Gln Tyr Leu Glu Glu 100 105 110Thr Lys Thr Phe
Ser Pro Gln Glu Ile Ser Ala Met Val Leu Thr Lys 115 120 125Met Lys
Glu Ile Ala Glu Ala Lys Ile Gly Lys Lys Val Glu Lys Ala 130 135
140Val Ile Thr Val Pro Ala Tyr Phe Asn Asp Ala Gln Arg Gln Ala
Thr145 150 155 160Lys Asp Ala Gly Ala Ile Ser Gly Leu Asn Val Leu
Arg Ile Ile Asn 165 170 175Glu Pro Thr Ala Ala Ala Ile Ala Tyr Gly
Leu Gly Ala Gly Lys Ser 180 185 190Glu Lys Glu Arg His Val Leu Ile
Phe Asp Leu Gly Gly Gly Thr Phe 195 200 205Asp Val Ser Leu Leu His
Ile Ala Gly Gly Val Tyr Thr Val Lys Ser 210 215 220Thr Ser Gly Asn
Thr His Leu Gly Gly Gln Asp Phe Asp Thr Asn Leu225 230 235 240Leu
Glu His Phe Lys Ala Glu Phe Lys Lys Lys Thr Gly Leu Asp Ile 245 250
255Ser Asp Asp Ala Arg Ala Leu Arg Arg Leu Arg Thr Ala Ala Glu Arg
260 265 270Ala Lys Arg Thr Leu Ser Ser Val Thr Gln Thr Thr Val Glu
Val Asp 275 280 285Ser Leu Phe Asp Gly Glu Asp Phe Glu Ser Ser Leu
Thr Arg Ala Arg 290 295 300Phe Glu Asp Leu Asn Ala Ala Leu Phe Lys
Ser Thr Leu Glu Pro Val305 310 315 320Glu Gln Val Leu Lys Asp Ala
Lys Ile Ser Lys Ser Gln Ile Asp Glu 325 330 335Val Val Leu Val Gly
Gly Ser Thr Arg Ile Pro Lys Val Gln Lys Leu 340 345 350Leu Ser Asp
Phe Phe Asp Gly Lys Gln Leu Glu Lys Ser Ile Asn Pro 355 360 365Asp
Glu Ala Val Ala Tyr Gly Ala Ala Val Gln Gly Ala Ile Leu Thr 370 375
380Gly Gln Ser Thr Ser Asp Glu Thr Lys Asp Leu Leu Leu Leu Asp
Val385 390 395 400Ala Pro Leu Ser Leu Gly Val Gly Met Gln Gly Asp
Ile Phe Gly Ile 405 410 415Val Val Pro Arg Asn Thr Thr Val Pro Thr
Ile Lys Arg Arg Thr Phe 420 425 430Thr Thr Val Ser Asp Asn Gln Thr
Thr Val Gln Phe Pro Val Tyr Gln 435 440 445Gly Glu Arg Val Asn Cys
Lys Glu Asn Thr Leu Leu Gly Glu Phe Asp 450 455 460Leu Lys Asn Ile
Pro Met Met Pro Ala Gly Glu Pro Val Leu Glu Ala465 470 475 480Ile
Phe Glu Val Asp Ala Asn Gly Ile Leu Lys Val Thr Ala Val Glu 485 490
495Lys Ser Thr Gly Lys Ser Ser Asn Ile Thr Ile Ser Asn Ala Val Gly
500 505 510Arg Leu Ser Ser Glu Glu Ile Glu Lys Met Val Asn Gln Ala
Glu Glu 515 520 525Phe Lys Ala Ala Asp Glu Ala Phe Ala Lys Lys His
Glu Ala Arg Gln 530 535 540Arg Leu Glu Ser Tyr Val Ala Ser Ile Glu
Gln Thr Val Thr Asp Pro545 550 555 560Val Leu Ser Ser Lys Leu Lys
Arg Gly Ser Lys Ser Lys Ile Glu Ala 565 570 575Ala Leu Ser Asp Ala
Leu Ala Ala Leu Gln Ile Glu Asp Pro Ser Ala 580 585 590Asp Glu Leu
Arg Lys Ala Glu Val Gly Leu Lys Arg Val Val Thr Lys 595 600 605Ala
Met Ser Ser Arg 610281842DNASaccharomyces cerevisiae 28atggctgaag
gtgttttcca aggtgctatc ggtatcgatt taggtacaac atactcttgt 60gttgctactt
atgaatcttc cgttgaaatt attgccaacg aacaaggtaa cagagttact
120ccatctttcg ttgccttcac cccacaggaa agattgatcg gtgatgctgc
caagaaccaa 180gctgctttga acccaagaaa cactgttttt gatgctaagc
gtttgattgg tagaagattc
240gacgacgagt ctgtccaaaa ggacatgaag acctggcctt tcaaggttat
cgacgtcgat 300ggtaacccag tcattgaagt ccaatacttg gaagaaacca
agactttctc cccacaagaa 360atttccgcta tggtcttgac caagatgaag
gaaattgctg aagctaagat tggtaagaag 420gttgaaaagg ctgtcattac
tgtcccagct tactttaacg atgcccaaag acaagctacc 480aaggatgccg
gtgccatttc tggtttgaac gttttgcgta tcatcaacga acctactgcc
540gctgctattg cttacggtct aggtgctggt aagtccgaaa aggaaagaca
tgttttgatt 600ttcgatttgg gtggtggtac tttcgatgtt tccttgttgc
acattgctgg tggtgtttac 660actgttaaat ctacttccgg taacactcac
ttgggtggtc aagatttcga caccaacttg 720ttggaacact tcaaggctga
attcaagaag aagactggtt tggacatctc cgacgatgcc 780agagctttga
gaagattgag aactgctgct gaaagagcta agagaacctt atcttctgtc
840actcaaacta ccgttgaagt tgactctttg tttgacggtg aagatttcga
atcctctttg 900actagagcta gatttgaaga cttgaacgcc gcattgttca
agtctacttt ggaacctgtt 960gaacaagttt tgaaggatgc taagatctct
aagtctcaaa tcgacgaagt tgtcttggtt 1020ggtggttcta ccagaattcc
aaaggtccaa aagttgttgt ctgacttctt tgacggtaag 1080caattggaaa
aatctattaa cccagatgaa gctgttgctt acggtgctgc tgttcaaggt
1140gctatcttga ctggccaatc cacatctgac gaaaccaagg acttgttgtt
gttagatgtt 1200gctccattat ctctaggtgt tggtatgcaa ggtgacattt
tcggtattgt tgtcccaaga 1260aacacaactg ttccaaccat caagagaaga
accttcacaa ctgtcagtga caaccaaacc 1320accgttcaat tcccagtcta
ccaaggtgaa cgtgtcaact gtaaagaaaa cactttgttg 1380ggtgaattcg
acttgaagaa catcccaatg atgccagctg gtgaaccagt cttggaagct
1440atcttcgaag ttgatgctaa cggtatcttg aaggttactg ccgtcgaaaa
gtctaccggt 1500aagtcttcta acatcactat ctccaacgct gtcggtagat
tgtcttctga agaaattgaa 1560aagatggtta accaagccga agagttcaag
gctgctgatg aagcttttgc taagaagcac 1620gaagctagac aaagactaga
atcctacgtc gcttccatcg aacaaaccgt cactgaccca 1680gtcttgtctt
ctaaattgaa gagaggttcc aagtccaaga tcgaagctgc tttgtccgat
1740gctttggctg ctttgcaaat cgaagaccca tccgctgatg agttgagaaa
ggcagaagtt 1800ggtttgaaga gagttgtcac caaggccatg tcttctcgtt aa
184229644PRTSaccharomyces cerevisiae 29Met Leu Pro Ser Trp Lys Ala
Phe Lys Ala His Asn Ile Leu Arg Ile1 5 10 15Leu Thr Arg Phe Gln Ser
Thr Lys Ile Pro Asp Ala Val Ile Gly Ile 20 25 30Asp Leu Gly Thr Thr
Asn Ser Ala Val Ala Ile Met Glu Gly Lys Val 35 40 45Pro Arg Ile Ile
Glu Asn Ala Glu Gly Ser Arg Thr Thr Pro Ser Val 50 55 60Val Ala Phe
Thr Lys Asp Gly Glu Arg Leu Val Gly Glu Pro Ala Lys65 70 75 80Arg
Gln Ser Val Ile Asn Ser Glu Asn Thr Leu Phe Ala Thr Lys Arg 85 90
95Leu Ile Gly Arg Arg Phe Glu Asp Ala Glu Val Gln Arg Asp Ile Asn
100 105 110Gln Val Pro Phe Lys Ile Val Lys His Ser Asn Gly Asp Ala
Trp Val 115 120 125Glu Ala Arg Asn Arg Thr Tyr Ser Pro Ala Gln Ile
Gly Gly Phe Ile 130 135 140Leu Asn Lys Met Lys Glu Thr Ala Glu Ala
Tyr Leu Ala Lys Ser Val145 150 155 160Lys Asn Ala Val Val Thr Val
Pro Ala Tyr Phe Asn Asp Ala Gln Arg 165 170 175Gln Ala Thr Lys Asp
Ala Gly Gln Ile Ile Gly Leu Asn Val Leu Arg 180 185 190Val Val Asn
Glu Pro Thr Ala Ala Ala Leu Ala Tyr Gly Leu Asp Lys 195 200 205Ser
Glu Pro Lys Val Ile Ala Val Phe Asp Leu Gly Gly Gly Thr Phe 210 215
220Asp Ile Ser Ile Leu Asp Ile Asp Asn Gly Ile Phe Glu Val Lys
Ser225 230 235 240Thr Asn Gly Asp Thr His Leu Gly Gly Glu Asp Phe
Asp Ile Tyr Leu 245 250 255Leu Gln Glu Ile Ile Ser His Phe Lys Lys
Glu Thr Gly Ile Asp Leu 260 265 270Ser Asn Asp Arg Met Ala Val Gln
Arg Ile Arg Glu Ala Ala Glu Lys 275 280 285Ala Lys Ile Glu Leu Ser
Ser Thr Leu Ser Thr Glu Ile Asn Leu Pro 290 295 300Phe Ile Thr Ala
Asp Ala Ala Gly Pro Lys His Ile Arg Met Pro Phe305 310 315 320Ser
Arg Val Gln Leu Glu Asn Ile Thr Ala Pro Leu Ile Asp Arg Thr 325 330
335Val Asp Pro Val Lys Lys Ala Leu Lys Asp Ala Arg Ile Thr Ala Ser
340 345 350Asp Ile Ser Asp Val Leu Leu Val Gly Gly Met Ser Arg Met
Pro Lys 355 360 365Val Ala Asp Thr Val Lys Lys Leu Phe Gly Lys Asp
Ala Ser Lys Ala 370 375 380Val Asn Pro Asp Glu Ala Val Ala Leu Gly
Ala Ala Ile Gln Ala Ala385 390 395 400Val Leu Ser Gly Glu Val Thr
Asp Val Leu Leu Leu Asp Val Thr Pro 405 410 415Leu Ser Leu Gly Ile
Glu Thr Leu Gly Gly Val Phe Thr Lys Leu Ile 420 425 430Pro Arg Asn
Ser Thr Ile Pro Asn Lys Lys Ser Gln Ile Phe Ser Thr 435 440 445Ala
Ala Ser Gly Gln Thr Ser Val Glu Val Lys Val Phe Gln Gly Glu 450 455
460Arg Glu Leu Val Lys Asp Asn Lys Leu Ile Gly Asn Phe Thr Leu
Ala465 470 475 480Gly Ile Pro Pro Ala Pro Lys Gly Thr Pro Gln Ile
Glu Val Thr Phe 485 490 495Asp Ile Asp Ala Asn Gly Ile Ile Asn Val
Ser Ala Lys Asp Leu Ala 500 505 510Ser His Lys Asp Ser Ser Ile Thr
Val Ala Gly Ala Ser Gly Leu Ser 515 520 525Asp Thr Glu Ile Asp Arg
Met Val Asn Glu Ala Glu Arg Tyr Lys Asn 530 535 540Gln Asp Arg Ala
Arg Arg Asn Ala Ile Glu Thr Ala Asn Lys Ala Asp545 550 555 560Gln
Leu Ala Asn Asp Thr Glu Asn Ser Ile Lys Glu Phe Glu Gly Lys 565 570
575Leu Asp Lys Thr Asp Ser Gln Arg Leu Lys Asp Gln Ile Ser Ser Leu
580 585 590Arg Glu Leu Val Ser Arg Ser Gln Ala Gly Asp Glu Val Asn
Asp Asp 595 600 605Asp Val Gly Thr Lys Ile Asp Asn Leu Arg Thr Ser
Ser Met Lys Leu 610 615 620Phe Glu Gln Leu Tyr Lys Asn Ser Asp Asn
Pro Glu Thr Lys Asn Gly625 630 635 640Arg Glu Asn
Lys301935DNASaccharomyces cerevisiae 30atgttaccat catggaaagc
ctttaaagca cataatatac ttcgtattct gacccgtttc 60cagtcaacca aaattccaga
tgcagttatc ggtattgatt taggtactac caattctgcg 120gtagctatta
tggaaggtaa agttccgaga attatcgaaa atgcagaagg ctcaagaact
180actccgtctg tagtggcttt cactaaagac ggagaacgtt tagttggtga
gccagccaaa 240cgacaatccg tcataaactc agaaaacact ttgtttgcta
ctaagcgttt aatcggccgc 300cgtttcgagg acgctgaagt ccaaagagat
attaatcagg ttcctttcaa aatcgtcaag 360cattctaatg gagatgcctg
ggtagaggct agaaacagaa cgtactcccc cgcccaaata 420ggaggtttta
tcttaaataa aatgaaggaa acagcggagg cttacttagc gaagagcgtc
480aaaaatgctg ttgtcaccgt tcctgcttac ttcaatgatg cccaaagaca
agctactaaa 540gacgcaggac aaattattgg gcttaatgta ttacgtgttg
tcaacgaacc aacagctgct 600gccctagctt acggtctaga taaatcagag
ccaaaagtca ttgctgtttt cgacttgggc 660ggtggtactt tcgatatttc
aatcctggac atcgataacg gtatctttga ggttaaatct 720accaatggtg
acacccattt gggtggcgaa gattttgaca tttatttgtt gcaagaaatt
780atttctcatt tcaagaaaga aaccggtatc gatttgagta atgaccgtat
ggctgtccaa 840agaataagag aagccgctga aaaggctaaa atcgaactgt
cttctacact ctctacagaa 900ataaacttgc ctttcataac tgctgatgct
gcaggcccaa agcatattcg tatgcccttt 960tctagggttc agcttgagaa
tataaccgcc ccattgattg atagaacggt tgatcctgtc 1020aaaaaagcac
tgaaagacgc aagaattacc gcctcagata tatcggatgt tttattagtt
1080ggtggtatgt caaggatgcc caaggttgca gatactgtaa agaaattatt
cggtaaggat 1140gcatcaaaag ctgttaaccc tgatgaagca gtcgctttag
gggccgctat acaggctgcg 1200gtcttgtctg gtgaagttac cgatgttttg
ttgctagatg tcactcccct atcattgggt 1260attgaaactt taggaggagt
ttttacaaaa ttaatcccaa gaaattctac aattcccaat 1320aagaaatctc
aaattttttc aactgcggca tcaggtcaaa catcggtgga agttaaagtt
1380ttccaaggtg agagggagtt agtcaaggat aacaaattaa taggtaattt
tactcttgcg 1440ggcattcctc cagctccaaa aggtacccca caaattgaag
tcacttttga tatcgatgcg 1500aacggcatca tcaacgtttc agcaaaagat
ctcgccagcc acaaagactc ttccatcact 1560gttgccggag cgtctgggct
atctgatacg gagattgatc gaatggttaa tgaagcggaa 1620agatataaaa
atcaggatag agccagaagg aatgccatcg aaaccgctaa caaagctgac
1680cagctagcta atgacacaga aaattccatt aaggaattcg aaggtaagct
agataaaact 1740gattctcaaa gactaaaaga tcaaatttca tccttaaggg
aattggtttc tcggagtcaa 1800gctggagatg aggttaatga tgacgatgtt
ggaacaaaaa ttgacaattt gcgaacttca 1860tcgatgaaac tttttgaaca
gttatacaag aacagtgaca atcctgaaac taagaacggg 1920agagaaaata aataa
193531511PRTSaccharomyces cerevisiae 31Met Ala Phe Gln Gln Gly Val
Leu Ser Arg Cys Ser Gly Val Phe Arg1 5 10 15His His Val Gly His Ser
Arg His Ile Asn Asn Ile Leu Tyr Arg His 20 25 30Ala Ile Ala Phe Ala
Ser Ile Ala Pro Arg Ile Pro Lys Ser Ser Phe 35 40 45His Thr Ser Ala
Ile Arg Asn Asn Glu Ala Phe Lys Asp Pro Tyr Asp 50 55 60Thr Leu Gly
Leu Lys Lys Ser Ala Thr Gly Ala Glu Ile Lys Lys Ala65 70 75 80Tyr
Tyr Lys Leu Ala Lys Lys Tyr His Pro Asp Ile Asn Lys Glu Pro 85 90
95Asp Ala Glu Lys Lys Phe His Asp Leu Gln Asn Ala Tyr Glu Ile Leu
100 105 110Ser Asp Glu Thr Lys Arg Gln Gln Tyr Asp Gln Phe Gly Pro
Ala Ala 115 120 125Phe Gly Gly Gly Gly Ala Ala Gly Gly Ala Gly Gly
Gly Ser Gly Ser 130 135 140Pro Phe Gly Ser Gln Phe His Asp Phe Ser
Gly Phe Thr Ser Ala Gly145 150 155 160Gly Ser Pro Phe Gly Gly Ile
Asn Phe Glu Asp Leu Phe Gly Ala Ala 165 170 175Phe Gly Gly Gly Gly
Arg Gly Ser Gly Gly Ala Ser Arg Ser Ser Ser 180 185 190Met Phe Arg
Gln Tyr Arg Gly Asp Pro Ile Glu Ile Val His Lys Val 195 200 205Ser
Phe Lys Asp Ala Val Phe Gly Ser Lys Asn Val Gln Leu Arg Phe 210 215
220Ser Ala Leu Asp Pro Cys Ser Thr Cys Ser Gly Thr Gly Met Lys
Pro225 230 235 240Asn Thr His Lys Val Ser Cys Ser Thr Cys His Gly
Thr Gly Thr Thr 245 250 255Val His Ile Arg Gly Gly Phe Gln Met Met
Ser Thr Cys Pro Thr Cys 260 265 270Asn Gly Glu Gly Thr Met Lys Arg
Pro Gln Asp Asn Cys Thr Lys Cys 275 280 285His Gly Glu Gly Val Gln
Val Asn Arg Ala Lys Thr Ile Thr Val Asp 290 295 300Leu Pro His Gly
Leu Gln Asp Gly Asp Val Val Arg Ile Pro Gly Gln305 310 315 320Gly
Ser Tyr Pro Asp Ile Ala Val Glu Ala Asp Leu Lys Asp Ser Val 325 330
335Lys Leu Ser Arg Gly Asp Ile Leu Val Arg Ile Arg Val Asp Lys Asp
340 345 350Pro Asn Phe Ser Ile Lys Asn Lys Tyr Asp Ile Trp Tyr Asp
Lys Glu 355 360 365Ile Pro Ile Thr Thr Ala Ala Leu Gly Gly Thr Val
Thr Ile Pro Thr 370 375 380Val Glu Gly Gln Lys Ile Arg Ile Lys Val
Ala Pro Gly Thr Gln Tyr385 390 395 400Asn Gln Val Ile Ser Ile Pro
Asn Met Gly Val Pro Lys Thr Ser Thr 405 410 415Ile Arg Gly Asp Met
Lys Val Gln Tyr Lys Ile Val Val Lys Lys Pro 420 425 430Gln Ser Leu
Ala Glu Lys Cys Leu Trp Glu Ala Leu Ala Asp Val Thr 435 440 445Asn
Asp Asp Met Ala Lys Lys Thr Met Gln Pro Gly Thr Ala Ala Gly 450 455
460Thr Ala Ile Asn Glu Glu Ile Leu Lys Lys Gln Lys Gln Glu Glu
Glu465 470 475 480Lys His Ala Lys Lys Asp Asp Asp Asn Thr Leu Lys
Arg Leu Glu Asn 485 490 495Phe Ile Thr Asn Thr Phe Arg Lys Ile Lys
Gly Asp Lys Lys Asn 500 505 510321536DNASaccharomyces cerevisiae
32atggctttcc aacaaggtgt attgtcaagg tgttccggtg tctttagaca ccatgtggga
60cattctcgcc atatcaataa tattctttat agacatgcca tcgcgtttgc atccatcgct
120ccacgaatac caaaatctag cttccatact tctgcaatca gaaacaacga
agcattcaag 180gacccgtacg atactttagg cttgaagaaa tctgctacag
gtgcggaaat caaaaaagca 240tactacaaac tggcaaagaa gtaccacccg
gatatcaaca aggaaccgga tgctgagaag 300aaattccacg atttacagaa
cgcttatgaa attctgtcag acgaaacgaa gaggcagcag 360tacgatcaat
ttgggcccgc tgccttcggc ggcggcggtg ccgctggagg tgccggtggt
420ggtagtggct ctccctttgg ttcccaattt catgatttct caggattcac
cagtgcaggc 480ggctcgccat ttggcggtat caattttgaa gacctgtttg
gtgctgcatt tggtggtggt 540ggccgcggta gcggtggcgc aagcaggtcg
tcatctatgt tcagacaata taggggcgac 600ccaatcgaga ttgtccataa
agtgtctttc aaggacgcag tgtttgggtc caagaacgtt 660cagttaagat
tctctgcgct ggacccttgt agtacctgtt cagggacggg aatgaaacca
720aacacgcata aggtcagttg tagcacttgt cacggaacag gaaccactgt
tcacattagg 780ggcggatttc agatgatgtc gacttgtcct acttgcaacg
gtgaaggtac catgaaacgg 840cctcaggaca attgtaccaa gtgccatggt
gagggtgttc aggtcaacag ggcaaagaca 900attacggtgg acttgccaca
tggattacag gacggcgacg tggtcaggat ccctggccaa 960ggctcatacc
ctgacatcgc tgtagaggcg gacttgaaag attcagtcaa gttatcaaga
1020ggtgatattt tggtgagaat tcgtgtcgac aaggatccca acttttcgat
aaagaacaag 1080tacgatattt ggtacgacaa ggagattcct ataaccacag
ctgcacttgg tggtactgtc 1140actatcccca ctgtggaggg acaaaagatc
aggataaagg tcgctccagg gactcaatac 1200aatcaagtga tatccattcc
taacatgggt gttcctaaaa catcaaccat tcgcggtgat 1260atgaaagtcc
agtacaagat cgttgttaag aaaccgcaat cgctggcaga aaaatgcttg
1320tgggaggcac tggcagatgt caccaacgat gacatggcca agaaaaccat
gcaaccgggc 1380acagccgcgg gtacagccat taatgaagag atactgaaga
aacaaaaaca agaagaggaa 1440aaacacgcaa aaaaggatga cgacaacact
ttgaagagac tagaaaattt cattaccaac 1500acattcagga agatcaaagg
tgacaaaaaa aattaa 153633146PRTSaccharomyces cerevisiae 33Met Val
Leu Pro Ile Ile Ile Gly Leu Gly Val Thr Met Val Ala Leu1 5 10 15Ser
Val Lys Ser Gly Leu Asn Ala Trp Thr Val Tyr Lys Thr Leu Ser 20 25
30Pro Leu Thr Ile Ala Lys Leu Asn Asn Ile Arg Ile Glu Asn Pro Thr
35 40 45Ala Gly Tyr Arg Asp Ala Leu Lys Phe Lys Ser Ser Leu Ile Asp
Glu 50 55 60Glu Leu Lys Asn Arg Leu Asn Gln Tyr Gln Gly Gly Phe Ala
Pro Arg65 70 75 80Met Thr Glu Pro Glu Ala Leu Leu Ile Leu Asp Ile
Ser Ala Arg Glu 85 90 95Ile Asn His Leu Asp Glu Lys Leu Leu Lys Lys
Lys His Arg Lys Ala 100 105 110Met Val Arg Asn His Pro Asp Arg Gly
Gly Ser Pro Tyr Met Ala Ala 115 120 125Lys Ile Asn Glu Ala Lys Glu
Val Leu Glu Arg Ser Val Leu Leu Arg 130 135 140Lys
Arg14534441DNASaccharomyces cerevisiae 34atggttttgc ctataataat
tggtttgggc gtgacaatgg ttgctctaag tgtcaagtct 60ggtctcaatg catggaccgt
ctacaagacc ctgtcccctt taactattgc aaaactaaat 120aacattcgca
tagaaaaccc gacggcgggc taccgcgatg cacttaagtt caaaagctca
180ctgatagacg aagaactgaa aaatagatta aaccagtacc agggaggctt
tgcaccgcga 240atgacagagc ccgaagcctt gctcatcttg gatatctccg
ccagagagat taatcacttg 300gatgaaaaat tactgaaaaa aaagcacagg
aaggctatgg ttcgtaacca cccagacaga 360ggagggagtc cctacatggc
ggccaagata aatgaggcga aagaagttct cgaaagaagt 420gttttactaa
gaaagagata a 44135563PRTSaccharomyces cerevisiae 35Met Arg Leu Arg
Thr Ala Ile Ala Thr Leu Cys Leu Thr Ala Phe Thr1 5 10 15Ser Ala Thr
Ser Asn Asn Ser Tyr Ile Ala Thr Asp Gln Thr Gln Asn 20 25 30Ala Phe
Asn Asp Thr His Phe Cys Lys Val Asp Arg Asn Asp His Val 35 40 45Ser
Pro Ser Cys Asn Val Thr Phe Asn Glu Leu Asn Ala Ile Asn Glu 50 55
60Asn Ile Arg Asp Asp Leu Ser Ala Leu Leu Lys Ser Asp Phe Phe Lys65
70 75 80Tyr Phe Arg Leu Asp Leu Tyr Lys Gln Cys Ser Phe Trp Asp Ala
Asn 85 90 95Asp Gly Leu Cys Leu Asn Arg Ala Cys Ser Val Asp Val Val
Glu Asp 100 105 110Trp Asp Thr Leu Pro Glu Tyr Trp Gln Pro Glu Ile
Leu Gly Ser Phe 115 120 125Asn Asn Asp Thr Met Lys Glu Ala Asp Asp
Ser Asp Asp Glu Cys Lys 130 135 140Phe Leu Asp Gln Leu Cys Gln Thr
Ser Lys Lys Pro Val Asp Ile Glu145 150 155 160Asp Thr Ile Asn Tyr
Cys Asp Val Asn Asp Phe Asn Gly Lys Asn Ala 165 170 175Val Leu Ile
Asp Leu Thr Ala Asn Pro Glu Arg Phe Thr Gly Tyr Gly 180 185 190Gly
Lys Gln Ala Gly Gln Ile Trp Ser Thr Ile Tyr Gln Asp Asn Cys 195 200
205Phe Thr Ile Gly Glu Thr Gly Glu Ser Leu Ala Lys Asp Ala Phe Tyr
210
215 220Arg Leu Val Ser Gly Phe His Ala Ser Ile Gly Thr His Leu Ser
Lys225 230 235 240Glu Tyr Leu Asn Thr Lys Thr Gly Lys Trp Glu Pro
Asn Leu Asp Leu 245 250 255Phe Met Ala Arg Ile Gly Asn Phe Pro Asp
Arg Val Thr Asn Met Tyr 260 265 270Phe Asn Tyr Ala Val Val Ala Lys
Ala Leu Trp Lys Ile Gln Pro Tyr 275 280 285Leu Pro Glu Phe Ser Phe
Cys Asp Leu Val Asn Lys Glu Ile Lys Asn 290 295 300Lys Met Asp Asn
Val Ile Ser Gln Leu Asp Thr Lys Ile Phe Asn Glu305 310 315 320Asp
Leu Val Phe Ala Asn Asp Leu Ser Leu Thr Leu Lys Asp Glu Phe 325 330
335Arg Ser Arg Phe Lys Asn Val Thr Lys Ile Met Asp Cys Val Gln Cys
340 345 350Asp Arg Cys Arg Leu Trp Gly Lys Ile Gln Thr Thr Gly Tyr
Ala Thr 355 360 365Ala Leu Lys Ile Leu Phe Glu Ile Asn Asp Ala Asp
Glu Phe Thr Lys 370 375 380Gln His Ile Val Gly Lys Leu Thr Lys Tyr
Glu Leu Ile Ala Leu Leu385 390 395 400Gln Thr Phe Gly Arg Leu Ser
Glu Ser Ile Glu Ser Val Asn Met Phe 405 410 415Glu Lys Met Tyr Gly
Lys Arg Leu Asn Gly Ser Glu Asn Arg Leu Ser 420 425 430Ser Phe Phe
Gln Asn Asn Phe Phe Asn Ile Leu Lys Glu Ala Gly Lys 435 440 445Ser
Ile Arg Tyr Thr Ile Glu Asn Ile Asn Ser Thr Lys Glu Gly Lys 450 455
460Lys Lys Thr Asn Asn Ser Gln Ser His Val Phe Asp Asp Leu Lys
Met465 470 475 480Pro Lys Ala Glu Ile Val Pro Arg Pro Ser Asn Gly
Thr Val Asn Lys 485 490 495Trp Lys Lys Ala Trp Asn Thr Glu Val Asn
Asn Val Leu Glu Ala Phe 500 505 510Arg Phe Ile Tyr Arg Ser Tyr Leu
Asp Leu Pro Arg Asn Ile Trp Glu 515 520 525Leu Ser Leu Met Lys Val
Tyr Lys Phe Trp Asn Lys Phe Ile Gly Val 530 535 540Ala Asp Tyr Val
Ser Glu Glu Thr Arg Glu Pro Ile Ser Tyr Lys Leu545 550 555 560Asp
Ile Gln361692DNASaccharomyces cerevisiae 36atgagattaa gaaccgccat
tgccacactg tgcctcacgg cttttacatc tgcaacttca 60aacaatagct acatcgccac
cgaccaaaca caaaatgcct ttaatgacac tcacttttgt 120aaggtcgaca
ggaatgatca cgttagtccc agttgtaacg taacattcaa tgaattaaat
180gccataaatg aaaacattag agatgatctt tcggcgttat taaaatctga
tttcttcaaa 240tactttcggc tggatttata caagcaatgt tcattttggg
acgccaacga tggtctgtgc 300ttaaaccgcg cttgctctgt tgatgtcgta
gaggactggg atacactgcc tgagtactgg 360cagcctgaga tcttgggtag
tttcaataat gatacaatga aggaagcgga tgatagcgat 420gacgaatgta
agttcttaga tcaactatgt caaaccagta aaaaacctgt agatatcgaa
480gacaccatca actactgtga tgtaaatgac tttaacggta aaaacgccgt
tctgattgat 540ttaacagcaa atccggaacg atttacaggt tatggtggta
agcaagctgg tcaaatttgg 600tctactatct accaagacaa ctgttttaca
attggcgaaa ctggtgaatc attggccaaa 660gatgcatttt atagacttgt
atccggtttc catgcctcta tcggtactca cttatcaaag 720gaatatttga
acacgaaaac tggtaaatgg gagcccaatc tggatttgtt tatggcaaga
780atcgggaact ttcctgatag agtgacaaac atgtatttca attatgctgt
tgtagctaag 840gctctctgga aaattcaacc atatttacca gaattttcat
tctgtgatct agtcaataaa 900gaaatcaaaa acaaaatgga taacgttatt
tcccagctgg acacaaaaat ttttaacgaa 960gacttagttt ttgccaacga
cctaagtttg actttgaagg acgaattcag atctcgcttc 1020aagaatgtca
cgaagattat ggattgtgtg caatgtgata gatgtagatt gtggggcaaa
1080attcaaacta ccggttacgc aactgccttg aaaattttgt ttgaaatcaa
cgacgctgat 1140gaattcacca aacaacatat tgttggtaag ttaaccaaat
atgagttgat tgcactatta 1200cagactttcg gtagattatc tgaatctatt
gaatctgtta acatgttcga aaaaatgtac 1260gggaaaaggt taaacggttc
tgaaaacagg ttaagctcat tcttccaaaa taacttcttc 1320aacattttga
aggaggcagg caaatcgatt cgttacacca tagagaacat caattccact
1380aaagaaggaa agaaaaagac taacaattct caatcacatg tatttgatga
tttaaaaatg 1440cccaaagcag aaatagttcc aaggccctct aacggtacag
taaataaatg gaagaaagct 1500tggaatactg aagttaacaa cgttttagaa
gcattcagat ttatttatag aagctatttg 1560gatttaccca ggaacatctg
ggaattatct ttgatgaagg tatacaaatt ttggaataaa 1620ttcatcggtg
ttgctgatta cgttagtgag gagacacgag agcctatttc ctataagcta
1680gatatacaat aa 169237196PRTSaccharomyces cerevisiae 37Met Lys
Gln Ile Val Lys Arg Ser His Ala Ile Arg Ile Val Ala Ala1 5 10 15Leu
Gly Ile Ile Gly Leu Trp Met Phe Phe Ser Ser Asn Glu Leu Ser 20 25
30Ile Ala Thr Pro Gly Leu Ile Lys Ala Lys Ser Gly Ile Asp Glu Val
35 40 45Gln Gly Ala Ala Ala Glu Lys Asn Asp Ala Arg Leu Lys Glu Ile
Glu 50 55 60Lys Gln Thr Ile Met Pro Leu Met Gly Asp Asp Lys Val Lys
Lys Glu65 70 75 80Val Gly Arg Ala Ser Trp Lys Tyr Phe His Thr Leu
Leu Ala Arg Phe 85 90 95Pro Asp Glu Pro Thr Pro Glu Glu Arg Glu Lys
Leu His Thr Phe Ile 100 105 110Gly Leu Tyr Ala Glu Leu Tyr Pro Cys
Gly Glu Cys Ser Tyr His Phe 115 120 125Val Lys Leu Ile Glu Lys Tyr
Pro Val Gln Thr Ser Ser Arg Thr Ala 130 135 140Ala Ala Met Trp Gly
Cys His Ile His Asn Lys Val Asn Glu Tyr Leu145 150 155 160Lys Lys
Asp Ile Tyr Asp Cys Ala Thr Ile Leu Glu Asp Tyr Asp Cys 165 170
175Gly Cys Ser Asp Ser Asp Gly Lys Arg Val Ser Leu Glu Lys Glu Ala
180 185 190Lys Gln His Gly 19538591DNASaccharomyces cerevisiae
38atgaaacaga tagtcaaaag aagccatgcc atcagaatag ttgcagcatt aggaatcata
60ggcctgtgga tgtttttctc gtctaatgaa ctatccatcg ctacgccggg cctaatcaag
120gcgaagtctg gtatagatga agtgcaaggg gcggctgctg agaagaacga
cgctcggttg 180aaagagatcg agaagcaaac cattatgcca ttgatgggcg
atgacaaggt gaagaaggaa 240gtgggcaggg cgtcgtggaa gtacttccat
accctgctgg cccgttttcc ggacgagcct 300actcctgaag aaagagagaa
actgcacacg tttattgggt tgtatgcaga actctatcca 360tgcggggaat
gttcatatca ctttgtaaag ttgattgaga agtatcccgt acagacatct
420agcaggacgg ctgccgcaat gtggggatgc cacattcaca acaaggtgaa
cgaataccta 480aagaaagaca tatatgactg tgctaccatc ctggaggact
acgattgtgg atgtagtgac 540agcgacggta aacgcgtgtc tctcgagaag
gaggctaaac agcacggttg a 59139517PRTSaccharomyces cerevisiae 39Met
Gln Val Thr Thr Arg Phe Ile Ser Ala Ile Val Ser Phe Cys Leu1 5 10
15Phe Ala Ser Phe Thr Leu Ala Glu Asn Ser Ala Arg Ala Thr Pro Gly
20 25 30Ser Asp Leu Leu Val Leu Thr Glu Lys Lys Phe Lys Ser Phe Ile
Glu 35 40 45Ser His Pro Leu Val Leu Val Glu Phe Phe Ala Pro Trp Cys
Leu His 50 55 60Ser Gln Ile Leu Arg Pro His Leu Glu Glu Ala Ala Ser
Ile Leu Lys65 70 75 80Glu His Asn Val Pro Val Val Gln Ile Asp Cys
Glu Ala Asn Ser Met 85 90 95Val Cys Leu Gln Gln Thr Ile Asn Thr Tyr
Pro Thr Leu Lys Ile Phe 100 105 110Lys Asn Gly Arg Ile Phe Asp Gly
Gln Val Tyr Arg Gly Val Lys Ile 115 120 125Thr Asp Glu Ile Thr Gln
Tyr Met Ile Gln Leu Tyr Glu Ala Ser Val 130 135 140Ile Tyr Leu Asn
Ser Glu Asp Glu Ile Gln Pro Tyr Leu Glu Asn Ala145 150 155 160Thr
Leu Pro Val Val Ile Asn Arg Gly Leu Thr Gly Leu Asn Glu Thr 165 170
175Tyr Gln Glu Val Ala Leu Asp Leu Ala Glu Asp Tyr Val Phe Leu Ser
180 185 190Leu Leu Asp Ser Glu Asp Lys Ser Leu Ser Ile His Leu Pro
Asn Thr 195 200 205Thr Glu Pro Ile Leu Phe Asp Gly Asn Val Asp Ser
Leu Val Gly Asn 210 215 220Ser Val Ala Leu Thr Gln Trp Leu Lys Val
Val Ile Leu Pro Tyr Phe225 230 235 240Thr Asp Ile Glu Pro Asp Leu
Phe Pro Lys Tyr Ile Ser Ser Asn Leu 245 250 255Pro Leu Ala Tyr Phe
Phe Tyr Thr Ser Glu Glu Glu Leu Glu Asp Tyr 260 265 270Thr Asp Leu
Phe Thr Gln Leu Gly Lys Glu Asn Arg Gly Gln Ile Asn 275 280 285Phe
Ile Ala Leu Asn Ser Thr Met Phe Pro His His Val Arg Phe Leu 290 295
300Asn Met Arg Glu Gln Phe Pro Leu Phe Ala Ile His Asn Met Ile
Asn305 310 315 320Asn Leu Lys Tyr Gly Leu Pro Gln Leu Pro Glu Glu
Glu Tyr Ala Lys 325 330 335Leu Glu Lys Pro Gln Pro Leu Asp Arg Asp
Met Ile Val Gln Leu Val 340 345 350Lys Asp Tyr Arg Glu Gly Thr Ala
Lys Pro Ile Val Lys Ser Glu Glu 355 360 365Ile Pro Lys Glu Gln Lys
Ser Asn Val Tyr Lys Ile Val Gly Lys Thr 370 375 380His Asp Asp Ile
Val His Asp Asp Asp Lys Asp Val Leu Val Lys Tyr385 390 395 400Tyr
Ala Thr Trp Cys Ile His Ser Lys Arg Phe Ala Pro Ile Tyr Glu 405 410
415Glu Ile Ala Asn Val Leu Ala Ser Asp Glu Ser Val Arg Asp Lys Ile
420 425 430Leu Ile Ala Glu Val Asp Ser Gly Ala Asn Asp Ile Leu Ser
Phe Pro 435 440 445Val Thr Gly Tyr Pro Thr Ile Ala Leu Tyr Pro Ala
Gly Asn Asn Ser 450 455 460Lys Pro Ile Ile Phe Asn Lys Ile Arg Asn
Leu Glu Asp Val Phe Glu465 470 475 480Phe Ile Lys Glu Ser Gly Thr
His His Ile Asp Gly Gln Ala Ile Tyr 485 490 495Asp Lys Leu His Gln
Ala Lys Asp Ser Glu Val Ser Thr Glu Asp Thr 500 505 510Val His Asp
Glu Leu 515401554DNASaccharomyces cerevisiae 40atgcaagtga
ccacaagatt tatatctgcg atagtctcgt tttgcctgtt tgcttctttc 60acgttggctg
aaaacagcgc aagagctacg ccgggatcag atttactcgt tctaacagag
120aagaaattta aatcattcat cgaatctcat ccgttagtcc tcgtcgagtt
ttttgctcca 180tggtgtttgc attctcagat cttacgccct cacttagaag
aggccgcctc tattttaaag 240gagcataacg tcccagttgt tcaaattgat
tgtgaggcta acagtatggt ttgcctgcaa 300caaactataa atacctaccc
aaccttgaaa atctttaaaa atggtcgtat ttttgatggt 360caagtctatc
gcggtgtcaa gatcaccgat gaaatcactc agtacatgat tcagctatac
420gaggcttctg tcatttattt aaattccgaa gatgaaatcc aaccatactt
ggaaaatgca 480actttaccag tagtaataaa cagaggcttg acaggcttga
atgaaacgta tcaagaagtc 540gcactggacc ttgctgagga ttacgtcttt
ttatcccttc tagattcaga agataagtca 600ttatcaatcc acttgccaaa
cactacagaa ccaattctgt ttgatggaaa tgtagactct 660ttggtcggaa
attccgttgc tctaactcag tggttaaaag tggtaatttt accttacttt
720accgacatcg aacctgatct cttccccaag tacatttcta gcaatttgcc
gttggcttac 780ttcttttata cttctgagga agaattggaa gattacactg
atcttttcac gcagttaggt 840aaggaaaatc gtggccaaat aaatttcatt
gcattaaact ctacaatgtt cccacaccac 900gttagattcc taaatatgag
agaacagttc ccattatttg ctatccataa tatgatcaat 960aatctgaaat
atggtttacc acaactacca gaagaagagt acgcgaaatt agaaaaacca
1020caaccactag acagagatat gatcgttcag ttggtaaaag attaccgtga
aggtactgcc 1080aagccaattg ttaagtcaga agagattcca aaagaacaaa
agtccaatgt ttataaaata 1140gttgggaaga cacatgacga cattgttcat
gatgatgaca aggatgtcct tgtcaaatat 1200tacgcgacat ggtgtattca
tagtaaaagg tttgcgccta tttacgaaga aattgcaaat 1260gtcttagcat
ctgatgaatc tgttcgcgat aaaatcttga tcgccgaagt agattcaggg
1320gcaaatgata tcttaagttt tcctgtgaca ggatatccaa ccattgcttt
gtatcctgcc 1380ggaaataact ctaagcctat tatcttcaat aaaattagaa
atttggaaga tgttttcgaa 1440tttatcaagg aatcaggtac acatcacatt
gacggccagg caatttatga taaattgcac 1500caggccaagg attctgaagt
gtctactgaa gataccgtac atgatgaatt ataa 155441318PRTSaccharomyces
cerevisiae 41Met Leu Phe Leu Asn Ile Ile Lys Leu Leu Leu Gly Leu
Phe Ile Met1 5 10 15Asn Glu Val Lys Ala Gln Asn Phe Tyr Asp Ser Asp
Pro His Ile Ser 20 25 30Glu Leu Thr Pro Lys Ser Phe Asp Lys Ala Ile
His Asn Thr Asn Tyr 35 40 45Thr Ser Leu Val Glu Phe Tyr Ala Pro Trp
Cys Gly His Cys Lys Lys 50 55 60Leu Ser Ser Thr Phe Arg Lys Ala Ala
Lys Arg Leu Asp Gly Val Val65 70 75 80Gln Val Ala Ala Val Asn Cys
Asp Leu Asn Lys Asn Lys Ala Leu Cys 85 90 95Ala Lys Tyr Asp Val Asn
Gly Phe Pro Thr Leu Met Val Phe Arg Pro 100 105 110Pro Lys Ile Asp
Leu Ser Lys Pro Ile Asp Asn Ala Lys Lys Ser Phe 115 120 125Ser Ala
His Ala Asn Glu Val Tyr Ser Gly Ala Arg Thr Leu Ala Pro 130 135
140Ile Val Asp Phe Ser Leu Ser Arg Ile Arg Ser Tyr Val Lys Lys
Phe145 150 155 160Val Arg Ile Asp Thr Leu Gly Ser Leu Leu Arg Lys
Ser Pro Lys Leu 165 170 175Ser Val Val Leu Phe Ser Lys Gln Asp Lys
Ile Ser Pro Val Tyr Lys 180 185 190Ser Ile Ala Leu Asp Trp Leu Gly
Lys Phe Asp Phe Tyr Ser Ile Ser 195 200 205Asn Lys Lys Leu Lys Gln
Leu Thr Asp Met Asn Pro Thr Tyr Glu Lys 210 215 220Thr Pro Glu Ile
Phe Lys Tyr Leu Gln Lys Val Ile Pro Glu Gln Arg225 230 235 240Gln
Ser Asp Lys Ser Lys Leu Val Val Phe Asp Ala Asp Lys Asp Lys 245 250
255Phe Trp Glu Tyr Glu Gly Asn Ser Ile Asn Lys Asn Asp Ile Ser Lys
260 265 270Phe Leu Arg Asp Thr Phe Ser Ile Thr Pro Asn Glu Gly Pro
Phe Ser 275 280 285Arg Arg Ser Glu Tyr Ile Ala Tyr Leu Lys Thr Gly
Lys Lys Pro Ile 290 295 300Lys Lys Asn His Ser Ser Ser Gly Asn Lys
His Asp Glu Leu305 310 31542957DNASaccharomyces cerevisiae
42atgttatttc ttaatattat taagctcctt ttgggacttt ttattatgaa tgaagtaaag
60gcgcaaaact tttacgattc cgatcctcat atatcagagt taacgccaaa aagcttcgat
120aaagcgatcc ataacacaaa ttacacatca ttagtggaat tttatgctcc
gtggtgcggc 180cattgtaaga agctctctag tacgttccgc aaggcagcaa
aaagattgga tggtgtagtc 240caagttgctg ctgtaaactg tgaccttaac
aagaataagg ctttgtgtgc taaatacgac 300gtaaacggat ttcccacgtt
aatggtattt aggcccccaa aaattgacct atctaagcca 360atagataacg
ccaaaaaaag tttcagcgct catgccaatg aagtgtactc aggtgcaaga
420actctcgcgc ctattgttga tttttctctt tcaagaataa ggtcatatgt
caaaaagttt 480gtccgtatag atacacttgg ctctttactt agaaagtcac
ccaaactttc cgtggtgttg 540ttttccaaac aagacaaaat ttcaccggtt
tataaaagca ttgcccttga ttggttagga 600aagttcgatt tttattcaat
ttcaaacaaa aaactcaagc aactaaccga tatgaaccca 660acatatgaaa
aaactcctga gattttcaaa tatttgcaga aggtcattcc tgaacagcga
720cagagcgata aaagtaagct tgtcgttttt gatgcagaca aagataaatt
ttgggagtat 780gaagggaact caatcaacaa aaatgacatt tccaaatttc
tgcgggacac ttttagtatt 840acccccaatg agggtccttt tagtagacgt
tctgaatata ttgcttactt aaaaactggc 900aagaagccaa ttaaaaagaa
ccattcctcc tcaggaaaca agcacgacga attgtag 95743277PRTSaccharomyces
cerevisiae 43Met Lys Leu His Gly Phe Leu Phe Ser Val Leu Ser Thr
Cys Val Val1 5 10 15Ile Leu Pro Ala Leu Ala Tyr Ser Glu Ala Val Thr
Met Val Lys Ser 20 25 30Ile Glu Gln Tyr Phe Asp Ile Cys Asn Arg Asn
Asp Ser Tyr Thr Met 35 40 45Ile Lys Tyr Tyr Thr Ser Trp Cys Gln His
Cys Lys Thr Leu Ala Pro 50 55 60Val Tyr Glu Glu Leu Gly Glu Leu Tyr
Ala Lys Lys Ala Asn Lys Asp65 70 75 80Asp Thr Pro Ile Asn Phe Leu
Glu Val Asn Cys Glu Phe Phe Gly Pro 85 90 95Thr Leu Cys Thr Asp Leu
Pro Gly Phe Pro Ile Ile Glu Leu Val Lys 100 105 110Pro Arg Thr Lys
Pro Leu Val Leu Pro Lys Leu Asp Trp Ser Ser Met 115 120 125Lys Phe
His Glu Arg Leu Trp Gln Arg Ile Lys Thr Trp Phe Asn Asn 130 135
140Pro Lys Tyr Gln Leu Asp Thr Ser Arg Val Val Arg Phe Glu Gly
Ser145 150 155 160Arg Asn Leu Lys Ser Leu Ser Asn Phe Ile Asp Thr
Val Arg Ser Lys 165 170 175Asp Thr Glu Glu Arg Phe Ile Glu His Ile
Phe Asp Asp Ser Arg Asn 180 185 190Cys Asn Glu Glu Leu Arg Ser Gln
Gln Leu Leu Cys Lys Ala Gly Lys 195 200 205Glu Tyr Tyr Ser Asp Thr
Leu Ser Lys Leu Tyr Gly Asp Val Asn Gly 210 215 220Leu Glu Lys Glu
Arg Arg Arg Leu Glu Ala Leu Ile Lys Gln Asn Gly225 230 235 240Asp
Asp Leu Ser Lys Glu Val Lys Glu Lys Leu Lys Ile Ile Arg Leu 245 250
255Gln Leu Ser Leu Leu Ser His Ile Glu Asp
Gln Leu Glu Asp Thr Ser 260 265 270Ser His Asp Glu Leu
27544834DNASaccharomyces cerevisiae 44atgaaattgc acggcttttt
attttccgta ttatcaacat gcgtcgtcat tttaccagcg 60ttggcctaca gtgaagctgt
cacgatggtc aagtcgattg agcagtactt cgatatctgc 120aataggaatg
attcttacac aatgataaaa tactacactt cttggtgcca acattgtaaa
180actctggccc cagtatacga agagcttggt gagctatacg ccaaaaaagc
taataaagat 240gataccccaa ttaacttcct tgaagttaac tgtgaattct
tcgggccaac tttatgtacc 300gacttgcctg gatttccaat aattgaactg
gtcaaacctc gtactaagcc cttagttctt 360ccgaagctcg attggtcgtc
tatgaaattt catgaaagac tatggcaaag aatcaagacg 420tggttcaaca
atcctaagta ccaactggat acgtctaggg ttgttcgttt tgaagggagt
480aggaacctaa agagtttaag caactttatc gatactgtaa gaagtaaaga
tacagaagaa 540agattcatag aacatatttt cgatgattct aggaattgca
atgaagaatt acgttctcaa 600cagcttctgt gtaaagctgg taaagaatac
tactctgata ctttatctaa attatacggt 660gacgtgaatg ggctggaaaa
ggaaaggcga agactagaag ctttaattaa gcaaaatgga 720gatgacttga
gtaaagaagt taaagaaaaa ctgaaaatca ttcgtctaca attgagccta
780ttatcacaca tagaagacca gttagaagat accagtagtc atgacgagct ttga
83445701PRTSaccharomyces cerevisiae 45Met Lys Met Asn Leu Lys Arg
Leu Val Val Thr Phe Phe Ser Cys Ile1 5 10 15Thr Phe Leu Leu Lys Phe
Thr Ile Ala Ala Ala Glu Pro Pro Glu Gly 20 25 30Phe Pro Glu Pro Leu
Asn Pro Thr Asn Phe Lys Glu Glu Leu Ser Lys 35 40 45Gly Leu His Ile
Ile Asp Phe Tyr Ser Pro Tyr Cys Pro His Cys Lys 50 55 60His Leu Ala
Pro Val Trp Met Glu Thr Trp Glu Glu Phe Lys Glu Glu65 70 75 80Ser
Lys Thr Leu Asn Ile Thr Phe Ser Gln Val Asn Cys Ile Glu Ser 85 90
95Ala Asp Leu Cys Gly Asp Glu Asn Ile Glu Tyr Phe Pro Glu Ile Arg
100 105 110Leu Tyr Asn Pro Ser Gly Tyr Ile Lys Ser Phe Thr Glu Thr
Pro Arg 115 120 125Thr Lys Glu Ser Leu Ile Ala Phe Ala Arg Arg Glu
Ser Met Asp Pro 130 135 140Asn Asn Leu Asp Thr Asp Leu Asp Ser Ala
Lys Ser Glu Ser Gln Tyr145 150 155 160Leu Glu Gly Phe Asp Phe Leu
Glu Leu Ile Ala Gly Lys Ala Thr Arg 165 170 175Pro His Leu Val Ser
Phe Trp Pro Thr Lys Asp Met Lys Asn Ser Asp 180 185 190Asp Ser Leu
Glu Phe Lys Asn Cys Asp Lys Cys His Glu Phe Gln Arg 195 200 205Thr
Trp Lys Ile Ile Ser Arg Gln Leu Ala Val Asp Asp Ile Asn Thr 210 215
220Gly His Val Asn Cys Glu Ser Asn Pro Thr Ile Cys Glu Glu Leu
Gly225 230 235 240Phe Gly Asp Leu Val Lys Ile Thr Asn His Arg Ala
Asp Arg Glu Pro 245 250 255Lys Val Ala Leu Val Leu Pro Asn Lys Thr
Ser Asn Asn Leu Phe Asp 260 265 270Tyr Pro Asn Gly Tyr Ser Ala Lys
Ser Asp Gly Tyr Val Asp Phe Ala 275 280 285Arg Arg Thr Phe Thr Asn
Ser Lys Phe Pro Asn Ile Thr Glu Gly Glu 290 295 300Leu Glu Lys Lys
Ala Asn Arg Asp Ile Asp Phe Leu Gln Glu Arg Gly305 310 315 320Arg
Val Thr Asn Asn Asp Ile His Leu Val Phe Ser Tyr Asp Pro Glu 325 330
335Thr Val Val Ile Glu Asp Phe Asp Ile Leu Glu Tyr Leu Ile Glu Pro
340 345 350Leu Ser Lys Ile Pro Asn Ile Tyr Leu His Gln Ile Asp Lys
Asn Leu 355 360 365Ile Asn Leu Ser Arg Asn Leu Phe Gly Arg Met Tyr
Glu Lys Ile Asn 370 375 380Tyr Asp Ala Ser Gln Thr Gln Lys Val Phe
Asn Lys Glu Tyr Phe Thr385 390 395 400Met Asn Thr Val Thr Gln Leu
Pro Thr Phe Phe Met Phe Lys Asp Gly 405 410 415Asp Pro Ile Ser Tyr
Val Phe Pro Gly Tyr Ser Thr Thr Glu Met Arg 420 425 430Asn Ile Asp
Ala Ile Met Asp Trp Val Lys Lys Tyr Ser Asn Pro Leu 435 440 445Val
Thr Glu Val Asp Ser Ser Asn Leu Lys Lys Leu Ile Ser Phe Gln 450 455
460Thr Lys Ser Tyr Ser Asp Leu Ala Ile Gln Leu Ile Ser Ser Thr
Asp465 470 475 480His Lys His Ile Lys Gly Ser Asn Lys Leu Ile Lys
Asn Leu Leu Leu 485 490 495Ala Ser Trp Glu Tyr Glu His Ile Arg Met
Glu Asn Asn Phe Glu Glu 500 505 510Ile Asn Glu Arg Arg Ala Arg Lys
Ala Asp Gly Ile Lys Lys Ile Lys 515 520 525Glu Lys Lys Ala Pro Ala
Asn Lys Ile Val Asp Lys Met Arg Glu Glu 530 535 540Ile Pro His Met
Asp Gln Lys Lys Leu Leu Leu Gly Tyr Leu Asp Ile545 550 555 560Ser
Lys Glu Lys Asn Phe Phe Arg Lys Tyr Gly Ile Thr Gly Glu Tyr 565 570
575Lys Ile Gly Asp Val Ile Ile Ile Asp Lys Ser Asn Asn Tyr Tyr Tyr
580 585 590Asn Lys Asp Asn Phe Gly Asn Ser Leu Thr Ser Asn Asn Pro
Gln Leu 595 600 605Leu Arg Glu Ala Phe Val Ser Leu Asn Ile Pro Ser
Lys Ala Leu Tyr 610 615 620Ser Ser Lys Leu Lys Gly Arg Leu Ile Asn
Ser Pro Phe His Asn Val625 630 635 640Leu Ser Phe Leu Asp Ile Ile
His Gly Asn Gly Met Pro Gly Tyr Leu 645 650 655Ile Val Ile Val Leu
Phe Ile Ala Ile Leu Lys Gly Pro Ser Ile Tyr 660 665 670Arg Arg Tyr
Lys Val Arg Lys His Tyr Arg Ala Lys Arg Asn Ala Val 675 680 685Gly
Ile Leu Gly Asn Met Glu Lys Lys Lys Asn Gln Asp 690 695
700462106DNASaccharomyces cerevisiae 46atgaaaatga atctgaaaag
gctcgtagtt accttcttct catgcatcac ctttctgctg 60aaattcacta tagccgccgc
tgaaccacca gagggctttc cagagccctt aaatccaaca 120aacttcaaag
aagagctatc taaggggctg catattattg acttctatag tccatactgt
180ccgcactgca aacatttagc acctgtttgg atggaaacat gggaggagtt
taaagaggag 240agcaaaacac tgaacataac attttcacag gttaactgca
tcgagagcgc cgatttgtgt 300ggagatgaaa atattgaata cttccctgaa
attagacttt ataacccctc aggatacatc 360aaatcgttca ctgaaacacc
gaggaccaaa gaatcattaa ttgcatttgc acgcagggag 420tctatggacc
caaataacct cgatactgat ctggattctg ctaaaagtga gagccagtat
480ctcgaaggct ttgattttct cgagctgatc gctggtaagg cgactaggcc
acatttggtt 540tccttctggc caacaaaaga tatgaaaaat agcgatgatt
cactagaatt caaaaactgt 600gacaaatgcc atgaattcca aaggacttgg
aagatcattt caagacagtt agccgtggat 660gatatcaaca cgggccacgt
taattgcgaa tctaatccaa caatctgtga agaactgggc 720tttggcgact
tggtgaaaat aaccaaccac agagccgata gagaacccaa ggtagcatta
780gtcctaccca ataaaacctc aaataatttg ttcgactatc ccaatggcta
ctcagcgaag 840tcagatggct atgtagattt tgccaggagg acttttacaa
acagtaaatt tcccaatata 900acagaagggg agctcgaaaa aaaagcaaac
agagacattg attttctgca agaaagggga 960cgagtaacta ataatgatat
ccatttagtt ttttcatatg accccgaaac tgttgttatt 1020gaagattttg
acattttgga gtatttaatc gagcctttgt caaaaattcc aaacatatat
1080ttgcaccaaa ttgacaagaa tctaataaat ttgtcacgta atctttttgg
aagaatgtat 1140gaaaagatca actacgacgc cagccaaact caaaaggttt
ttaacaaaga atactttact 1200atgaatacgg ttacgcaact cccaactttt
ttcatgttta aagatggtga tcccatatcc 1260tatgttttcc ccggatactc
cacaacagaa atgagaaata ttgatgccat tatggattgg 1320gtaaaaaagt
attctaatcc cttagttacc gaagttgact cttctaattt gaaaaaatta
1380atttccttcc aaaccaagag ctacagtgat ttagcaattc agttaataag
tagcactgac 1440cacaaacata tcaaaggaag caacaagctt attaaaaact
tgctcctcgc aagttgggag 1500tatgaacata ttcggatgga aaataacttc
gaagaaatta atgagagaag ggcaaggaaa 1560gcagacggga tcaagaaaat
aaaggaaaaa aaggctccgg ctaacaaaat tgttgataaa 1620atgcgtgaag
agattcccca tatggatcaa aaaaaattgt tattaggata tttagatatt
1680tcaaaggaga agaatttttt tagaaaatat ggtattactg gagaatataa
aattggtgat 1740gtgattatca ttgataaatc aaataattac tactacaata
aagataattt tggcaactcc 1800ttgacttcta acaaccctca attgctgaga
gaagcattcg tgtccttaaa tattccatca 1860aaagctctat acagctctaa
gttgaagggg agattgataa attctccatt ccataatgtc 1920ctcagtttcc
tagacataat ccacgggaac ggcatgcccg gttacttaat tgttattgtt
1980ttgtttatcg caatactcaa aggtccatct atttacagaa gatacaaagt
aaggaaacac 2040tatagggcga aaaggaacgc tgtcggtatc ctaggaaata
tggagaaaaa aaaaaatcaa 2100gattaa 210647522PRTSaccharomyces
cerevisiae 47Met Lys Phe Ser Ala Gly Ala Val Leu Ser Trp Ser Ser
Leu Leu Leu1 5 10 15Ala Ser Ser Val Phe Ala Gln Gln Glu Ala Val Ala
Pro Glu Asp Ser 20 25 30Ala Val Val Lys Leu Ala Thr Asp Ser Phe Asn
Glu Tyr Ile Gln Ser 35 40 45His Asp Leu Val Leu Ala Glu Phe Phe Ala
Pro Trp Cys Gly His Cys 50 55 60Lys Asn Met Ala Pro Glu Tyr Val Lys
Ala Ala Glu Thr Leu Val Glu65 70 75 80Lys Asn Ile Thr Leu Ala Gln
Ile Asp Cys Thr Glu Asn Gln Asp Leu 85 90 95Cys Met Glu His Asn Ile
Pro Gly Phe Pro Ser Leu Lys Ile Phe Lys 100 105 110Asn Ser Asp Val
Asn Asn Ser Ile Asp Tyr Glu Gly Pro Arg Thr Ala 115 120 125Glu Ala
Ile Val Gln Phe Met Ile Lys Gln Ser Gln Pro Ala Val Ala 130 135
140Val Val Ala Asp Leu Pro Ala Tyr Leu Ala Asn Glu Thr Phe Val
Thr145 150 155 160Pro Val Ile Val Gln Ser Gly Lys Ile Asp Ala Asp
Phe Asn Ala Thr 165 170 175Phe Tyr Ser Met Ala Asn Lys His Phe Asn
Asp Tyr Asp Phe Val Ser 180 185 190Ala Glu Asn Ala Asp Asp Asp Phe
Lys Leu Ser Ile Tyr Leu Pro Ser 195 200 205Ala Met Asp Glu Pro Val
Val Tyr Asn Gly Lys Lys Ala Asp Ile Ala 210 215 220Asp Ala Asp Val
Phe Glu Lys Trp Leu Gln Val Glu Ala Leu Pro Tyr225 230 235 240Phe
Gly Glu Ile Asp Gly Ser Val Phe Ala Gln Tyr Val Glu Ser Gly 245 250
255Leu Pro Leu Gly Tyr Leu Phe Tyr Asn Asp Glu Glu Glu Leu Glu Glu
260 265 270Tyr Lys Pro Leu Phe Thr Glu Leu Ala Lys Lys Asn Arg Gly
Leu Met 275 280 285Asn Phe Val Ser Ile Asp Ala Arg Lys Phe Gly Arg
His Ala Gly Asn 290 295 300Leu Asn Met Lys Glu Gln Phe Pro Leu Phe
Ala Ile His Asp Met Thr305 310 315 320Glu Asp Leu Lys Tyr Gly Leu
Pro Gln Leu Ser Glu Glu Ala Phe Asp 325 330 335Glu Leu Ser Asp Lys
Ile Val Leu Glu Ser Lys Ala Ile Glu Ser Leu 340 345 350Val Lys Asp
Phe Leu Lys Gly Asp Ala Ser Pro Ile Val Lys Ser Gln 355 360 365Glu
Ile Phe Glu Asn Gln Asp Ser Ser Val Phe Gln Leu Val Gly Lys 370 375
380Asn His Asp Glu Ile Val Asn Asp Pro Lys Lys Asp Val Leu Val
Leu385 390 395 400Tyr Tyr Ala Pro Trp Cys Gly His Cys Lys Arg Leu
Ala Pro Thr Tyr 405 410 415Gln Glu Leu Ala Asp Thr Tyr Ala Asn Ala
Thr Ser Asp Val Leu Ile 420 425 430Ala Lys Leu Asp His Thr Glu Asn
Asp Val Arg Gly Val Val Ile Glu 435 440 445Gly Tyr Pro Thr Ile Val
Leu Tyr Pro Gly Gly Lys Lys Ser Glu Ser 450 455 460Val Val Tyr Gln
Gly Ser Arg Ser Leu Asp Ser Leu Phe Asp Phe Ile465 470 475 480Lys
Glu Asn Gly His Phe Asp Val Asp Gly Lys Ala Leu Tyr Glu Glu 485 490
495Ala Gln Glu Lys Ala Ala Glu Glu Ala Asp Ala Asp Ala Glu Leu Ala
500 505 510Asp Glu Glu Asp Ala Ile His Asp Glu Leu 515
52048530PRTSaccharomyces cerevisiae 48Met Lys Phe Ser Ala Gly Ala
Val Leu Ser Trp Ser Ser Leu Leu Leu1 5 10 15Ala Ser Ser Val Phe Ala
Gln Gln Glu Ala Val Ala Pro Glu Asp Ser 20 25 30Ala Val Val Lys Leu
Ala Thr Asp Ser Phe Asn Glu Tyr Ile Gln Ser 35 40 45His Asp Leu Val
Leu Ala Glu Phe Phe Ala Pro Trp Cys Gly His Cys 50 55 60Lys Asn Met
Ala Pro Glu Tyr Val Lys Ala Ala Glu Thr Leu Val Glu65 70 75 80Lys
Asn Ile Thr Leu Ala Gln Ile Asp Cys Thr Glu Asn Gln Asp Leu 85 90
95Cys Met Glu His Asn Ile Pro Gly Phe Pro Ser Leu Lys Ile Phe Lys
100 105 110Asn Arg Asp Val Asn Asn Ser Ile Asp Tyr Glu Gly Pro Arg
Thr Ala 115 120 125Glu Ala Ile Val Gln Phe Met Ile Lys Gln Ser Gln
Pro Ala Val Ala 130 135 140Val Val Ala Asp Leu Pro Ala Tyr Leu Ala
Asn Glu Thr Phe Val Thr145 150 155 160Pro Val Ile Val Gln Ser Gly
Lys Ile Asp Ala Asp Phe Asn Ala Thr 165 170 175Phe Tyr Ser Met Ala
Asn Lys His Phe Asn Asp Tyr Asp Phe Val Ser 180 185 190Ala Glu Asn
Ala Asp Asp Asp Phe Lys Leu Ser Ile Tyr Leu Pro Ser 195 200 205Ala
Met Asp Glu Pro Val Val Tyr Asn Gly Lys Lys Ala Asp Ile Ala 210 215
220Asp Ala Asp Val Phe Glu Lys Trp Leu Gln Val Glu Ala Leu Pro
Tyr225 230 235 240Phe Gly Glu Ile Asp Gly Ser Val Phe Ala Gln Tyr
Val Glu Ser Gly 245 250 255Leu Pro Leu Gly Tyr Leu Phe Tyr Asn Asp
Glu Glu Glu Leu Glu Glu 260 265 270Tyr Lys Pro Leu Phe Thr Glu Leu
Ala Lys Lys Asn Arg Gly Leu Met 275 280 285Asn Phe Val Ser Ile Asp
Ala Arg Lys Phe Gly Arg His Ala Gly Asn 290 295 300Leu Asn Met Lys
Glu Gln Phe Pro Leu Phe Ala Ile His Asp Met Thr305 310 315 320Glu
Asp Leu Lys Tyr Gly Leu Pro Gln Leu Ser Glu Glu Ala Phe Asp 325 330
335Glu Leu Ser Asp Lys Ile Val Leu Glu Ser Lys Ala Ile Glu Ser Leu
340 345 350Val Lys Asp Phe Leu Lys Gly Asp Ala Ser Pro Ile Val Lys
Ser Gln 355 360 365Glu Ile Phe Glu Asn Gln Asp Ser Ser Val Phe Gln
Leu Val Gly Lys 370 375 380Asn His Asp Glu Ile Val Asn Asp Pro Lys
Lys Asp Val Leu Val Leu385 390 395 400Tyr Tyr Ala Pro Trp Cys Gly
His Cys Lys Arg Leu Ala Pro Thr Tyr 405 410 415Gln Glu Leu Ala Asp
Thr Tyr Ala Asn Ala Thr Ser Asp Val Leu Ile 420 425 430Ala Lys Leu
Asp His Thr Glu Asn Asp Val Arg Gly Val Val Ile Glu 435 440 445Gly
Tyr Pro Thr Ile Val Leu Tyr Pro Gly Gly Lys Lys Ser Glu Ser 450 455
460Val Val Tyr Gln Gly Ser Arg Ser Leu Asp Ser Leu Phe Asp Phe
Ile465 470 475 480Lys Glu Asn Gly His Phe Asp Val Asp Gly Lys Ala
Leu Tyr Glu Glu 485 490 495Ala Gln Glu Lys Ala Ala Glu Glu Ala Glu
Ala Asp Ala Glu Ala Glu 500 505 510Ala Asp Ala Asp Ala Glu Leu Ala
Asp Glu Glu Asp Ala Ile His Asp 515 520 525Glu Leu
53049211PRTSaccharomyces cerevisiae 49Met Asp Ala Val Ile Leu Asn
Leu Leu Gly Asp Ile Pro Leu Val Thr1 5 10 15Arg Leu Trp Thr Ile Gly
Cys Leu Val Leu Ser Gly Leu Thr Ser Leu 20 25 30Arg Ile Val Asp Pro
Gly Lys Val Val Tyr Ser Tyr Asp Leu Val Phe 35 40 45Lys Lys Gly Gln
Tyr Gly Arg Leu Leu Tyr Ser Ile Phe Asp Tyr Gly 50 55 60Ala Phe Asn
Trp Ile Ser Met Ile Asn Ile Phe Val Ser Ala Asn His65 70 75 80Leu
Ser Thr Leu Glu Asn Ser Phe Asn Leu Arg Arg Lys Phe Cys Trp 85 90
95Ile Ile Phe Leu Leu Leu Val Ile Leu Val Lys Met Thr Ser Ile Glu
100 105 110Gln Pro Ala Ala Ser Leu Gly Val Leu Leu His Glu Asn Leu
Val Tyr 115 120 125Tyr Glu Leu Lys Lys Asn Gly Asn Gln Met Asn Val
Arg Phe Phe Gly 130 135 140Ala Ile Asp Val Ser Pro Ser Ile Phe Pro
Ile Tyr Met Asn Ala Val145 150 155 160Met Tyr Phe Val Tyr Lys Arg
Ser Trp Leu Glu Ile Ala Met Asn Phe 165 170 175Met Pro Gly His Val
Ile Tyr Tyr Met Asp Asp Ile
Ile Gly Lys Ile 180 185 190Tyr Gly Ile Asp Leu Cys Lys Ser Pro Tyr
Asp Trp Phe Arg Asn Thr 195 200 205Glu Thr Pro
21050636DNASaccharomyces cerevisiae 50atggatgctg taatactgaa
tctcttaggc gacattcctt tggtcacaag attatggaca 60attggctgtc ttgtactatc
aggtctcaca agtctccgga ttgtggatcc agggaaggta 120gtgtacagtt
atgatttagt attcaaaaag ggacaatatg gaagactact ttattcgata
180ttcgattacg gcgcatttaa ttggatatcc atgataaaca tctttgtcag
cgctaatcac 240ttatcaactt tggaaaactc attcaatctg agaagaaaat
tctgttggat aatattttta 300ctgttggtga tactggtaaa gatgaccagc
attgaacaac ctgcagcatc actcggtgtg 360ttattgcatg agaatctcgt
gtactacgaa ctgaaaaaga acggaaacca aatgaacgta 420cgattcttcg
gtgccattga tgtttcacca tctatattcc caatctacat gaatgcagta
480atgtattttg tatataagcg tagctggtta gaaattgcca tgaatttcat
gccaggtcac 540gtaatttact acatggatga tataataggg aagatttatg
gcatcgattt gtgtaaatct 600ccgtacgact ggttccgcaa cactgaaaca ccctaa
63651551PRTSaccharomyces cerevisiae 51Met Val Pro Glu Asn Arg Arg
Lys Gln Leu Ala Ile Phe Val Val Val1 5 10 15Thr Tyr Leu Leu Thr Phe
Tyr Cys Val Tyr Ser Ala Thr Lys Thr Ser 20 25 30Val Ser Phe Leu Gln
Val Thr Leu Lys Leu Asn Glu Gly Phe Asn Leu 35 40 45Met Val Leu Ser
Ile Phe Ile Leu Leu Asn Ser Thr Leu Leu Trp Gln 50 55 60Leu Leu Thr
Lys Leu Leu Phe Gly Glu Leu Arg Leu Ile Glu His Glu65 70 75 80His
Ile Phe Glu Arg Leu Pro Phe Thr Ile Ile Asn Thr Leu Phe Met 85 90
95Ser Ser Leu Phe His Glu Arg Tyr Phe Phe Thr Val Ala Phe Phe Gly
100 105 110Leu Leu Leu Leu Tyr Leu Lys Val Phe His Trp Ile Leu Lys
Asp Arg 115 120 125Leu Glu Ala Leu Leu Gln Ser Ile Asn Asp Ser Thr
Thr Met Lys Thr 130 135 140Leu Ile Phe Ser Arg Phe Ser Phe Asn Leu
Val Leu Leu Ala Val Val145 150 155 160Asp Tyr Gln Ile Ile Thr Arg
Cys Ile Ser Ser Ile Tyr Thr Asn Gln 165 170 175Lys Ser Asp Ile Glu
Ser Thr Ser Leu Tyr Leu Ile Gln Val Met Glu 180 185 190Phe Thr Met
Leu Leu Ile Asp Leu Leu Asn Leu Phe Leu Gln Thr Cys 195 200 205Leu
Asn Phe Trp Glu Phe Tyr Arg Ser Gln Gln Ser Leu Ser Asn Glu 210 215
220Asn Asn His Ile Val His Gly Asp Pro Thr Asp Glu Asn Thr Val
Glu225 230 235 240Ser Asp Gln Ser Gln Pro Val Leu Asn Asp Asp Asp
Asp Asp Asp Asp 245 250 255Asp Asp Arg Gln Phe Thr Gly Leu Glu Gly
Lys Phe Met Tyr Glu Lys 260 265 270Ala Ile Asp Val Phe Thr Arg Phe
Leu Lys Thr Ala Leu His Leu Ser 275 280 285Met Leu Ile Pro Phe Arg
Met Pro Met Met Leu Leu Lys Asp Val Val 290 295 300Trp Asp Ile Leu
Ala Leu Tyr Gln Ser Gly Thr Ser Leu Trp Lys Ile305 310 315 320Trp
Arg Asn Asn Lys Gln Leu Asp Asp Thr Leu Val Thr Val Thr Val 325 330
335Glu Gln Leu Gln Asn Ser Ala Asn Asp Asp Asn Ile Cys Ile Ile Cys
340 345 350Met Asp Glu Leu Ile His Ser Pro Asn Gln Gln Thr Trp Lys
Asn Lys 355 360 365Asn Lys Lys Pro Lys Arg Leu Pro Cys Gly His Ile
Leu His Leu Ser 370 375 380Cys Leu Lys Asn Trp Met Glu Arg Ser Gln
Thr Cys Pro Ile Cys Arg385 390 395 400Leu Pro Val Phe Asp Glu Lys
Gly Asn Val Val Gln Thr Thr Phe Thr 405 410 415Ser Asn Ser Asp Ile
Thr Thr Gln Thr Thr Val Thr Asp Ser Thr Gly 420 425 430Ile Ala Thr
Asp Gln Gln Gly Phe Ala Asn Glu Val Asp Leu Leu Pro 435 440 445Thr
Arg Thr Thr Ser Pro Asp Ile Arg Ile Val Pro Thr Gln Asn Ile 450 455
460Asp Thr Leu Ala Met Arg Thr Arg Ser Thr Ser Thr Pro Ser Pro
Thr465 470 475 480Trp Tyr Thr Phe Pro Leu His Lys Thr Gly Asp Asn
Ser Val Gly Ser 485 490 495Ser Arg Ser Ala Tyr Glu Phe Leu Ile Thr
Asn Ser Asp Glu Lys Glu 500 505 510Asn Gly Ile Pro Val Lys Leu Thr
Ile Glu Asn His Glu Val Asn Ser 515 520 525Leu His Gly Asp Gly Gly
Glu Gln Ile Ala Lys Lys Ile Val Ile Pro 530 535 540Asp Lys Phe Ile
Gln His Ile545 550521656DNASaccharomyces cerevisiae 52atggtgccag
aaaatagaag gaaacagttg gcaatttttg tagttgtcac atatttgctc 60acattttatt
gcgtgtattc agccaccaag acaagcgttt cctttttgca agtaacactg
120aagctaaatg aaggcttcaa tctaatggtt ttgtcgatat tcatcttatt
aaattctacc 180ttactatggc aactcctaac gaaactatta tttggtgaac
tgaggcttat tgagcatgag 240cacatttttg aaaggttacc atttaccatt
ataaacacct tgtttatgtc ctcactgttc 300cacgaacggt attttttcac
agtggcattt tttggactat tactactcta tctgaaagtt 360ttccattgga
ttttaaagga taggctggag gccttattac agtcaataaa tgattccacc
420acaatgaaaa cccttatctt tagtagattc tcatttaacc tcgtactatt
ggcggttgta 480gactaccaga taataacacg atgcatctcc tccatatata
caaaccaaaa gagtgatatt 540gaatccacat ccctttacct gatacaagta
atggagttta ccatgctttt gattgatttg 600ctaaatttat tcctacagac
ttgtttgaat ttctgggaat tttatcgctc acaacaaagt 660ctgtctaatg
agaacaacca tattgtccat ggcgatccta cagatgaaaa cacggttgag
720tctgatcaat ctcagccagt gctgaatgac gacgacgatg acgacgatga
tgatagacaa 780tttaccggcc tggagggtaa attcatgtat gaaaaagcaa
ttgacgtatt cacaagattc 840ttaaaaacgg cacttcattt gtctatgcta
ataccattta ggatgcctat gatgcttttg 900aaagatgtgg tgtgggatat
cttggcacta tatcaaagtg gcacaagttt gtggaaaatc 960tggagaaata
acaaacagct cgacgacact cttgtcactg tcaccgtaga acagctacaa
1020aattctgcaa atgatgacaa tatttgtatc atttgtatgg atgagttaat
acattctcca 1080aaccagcaga cgtggaagaa taaaaacaag aaacccaaaa
ggttaccttg tggccacata 1140cttcatttgt cgtgtttaaa gaattggatg
gaacgttctc agacttgtcc tatttgtaga 1200ttgcctgtct ttgatgaaaa
aggtaatgtt gtgcaaacga ctttcacttc caatagtgat 1260atcacgacac
agaccaccgt aacagatagc actgggatag cgacagatca acaaggtttc
1320gcaaacgaag tagatctact tcccacaaga acaacttccc ctgatataag
gatagtgcct 1380actcaaaata tagacacatt agcaatgaga acaaggtcaa
cctctacacc atctcctacg 1440tggtatacgt tcccattaca taaaactggt
gataattctg ttgggtcaag ccgatcagcc 1500tacgaatttt tgatcacaaa
ttcagatgag aaagaaaatg gtattcctgt caaattaaca 1560atagaaaatc
acgaagtaaa ttctctgcat ggagacgggg gcgagcaaat tgccaagaaa
1620attgtcatac cagataaatt tatccagcat atctag
165653833PRTSaccharomyces cerevisiae 53Met Ile Thr Leu Leu Leu Tyr
Leu Cys Val Ile Cys Asn Ala Ile Val1 5 10 15Leu Ile Arg Ala Asp Ser
Ile Ala Asp Pro Trp Pro Glu Ala Arg His 20 25 30Leu Leu Asn Thr Ile
Ala Lys Ser Arg Asp Pro Met Lys Glu Ala Ala 35 40 45Met Glu Pro Asn
Ala Asp Glu Phe Val Gly Phe Tyr Val Pro Met Asp 50 55 60Tyr Ser Pro
Arg Asn Glu Glu Lys Asn Tyr Gln Ser Ile Trp Gln Asn65 70 75 80Glu
Ile Thr Asp Ser Gln Arg His Ile Tyr Glu Leu Leu Val Gln Ser 85 90
95Ser Glu Gln Phe Asn Asn Ser Glu Ala Thr Tyr Thr Leu Ser Gln Ile
100 105 110His Leu Trp Ser Gln Tyr Asn Phe Pro His Asn Met Thr Leu
Ala His 115 120 125Lys Tyr Leu Glu Lys Phe Asn Asp Leu Thr His Phe
Thr Asn His Ser 130 135 140Ala Ile Phe Asp Leu Ala Val Met Tyr Ala
Thr Gly Gly Cys Ala Ser145 150 155 160Gly Asn Asp Gln Thr Val Ile
Pro Gln Asp Ser Ala Lys Ala Leu Leu 165 170 175Tyr Tyr Gln Arg Ala
Ala Gln Leu Gly Asn Leu Lys Ala Lys Gln Val 180 185 190Leu Ala Tyr
Lys Tyr Tyr Ser Gly Phe Asn Val Pro Arg Asn Phe His 195 200 205Lys
Ser Leu Val Leu Tyr Arg Asp Ile Ala Glu Gln Leu Arg Lys Ser 210 215
220Tyr Ser Arg Asp Glu Trp Asp Ile Val Phe Pro Tyr Trp Glu Ser
Tyr225 230 235 240Asn Val Arg Ile Ser Asp Phe Glu Ser Gly Leu Leu
Gly Lys Gly Leu 245 250 255Asn Ser Val Pro Ser Ser Thr Val Arg Lys
Arg Thr Thr Arg Pro Asp 260 265 270Ile Gly Ser Pro Phe Ile Ala Gln
Val Asn Gly Val Gln Met Thr Leu 275 280 285Gln Ile Glu Pro Met Gly
Arg Phe Ala Phe Asn Gly Asn Asp Gly Asn 290 295 300Ile Asn Gly Asp
Glu Asp Asp Glu Asp Ala Ser Glu Arg Arg Ile Ile305 310 315 320Arg
Ile Tyr Tyr Ala Ala Leu Asn Asp Tyr Lys Gly Thr Tyr Ser Gln 325 330
335Ser Arg Asn Cys Glu Arg Ala Lys Asn Leu Leu Glu Leu Thr Tyr Lys
340 345 350Glu Phe Gln Pro His Val Asp Asn Leu Asp Pro Leu Gln Val
Phe Tyr 355 360 365Tyr Val Arg Cys Leu Gln Leu Leu Gly His Met Tyr
Phe Thr Gly Glu 370 375 380Gly Ser Ser Lys Pro Asn Ile His Met Ala
Glu Glu Ile Leu Thr Thr385 390 395 400Ser Leu Glu Ile Ser Arg Arg
Ala Gln Gly Pro Ile Gly Arg Ala Cys 405 410 415Ile Asp Leu Gly Leu
Ile Asn Gln Tyr Ile Thr Asn Asn Ile Ser Gln 420 425 430Ala Ile Ser
Tyr Tyr Met Lys Ala Met Lys Thr Gln Ala Asn Asn Gly 435 440 445Ile
Val Glu Phe Gln Leu Ser Lys Leu Ala Thr Ser Phe Pro Glu Glu 450 455
460Lys Ile Gly Asp Pro Phe Asn Leu Met Glu Thr Ala Tyr Leu Asn
Gly465 470 475 480Phe Ile Pro Ala Ile Tyr Glu Phe Ala Val Met Ile
Glu Ser Gly Met 485 490 495Asn Ser Lys Ser Ser Val Glu Asn Thr Ala
Tyr Leu Phe Lys Thr Phe 500 505 510Val Asp Lys Asn Glu Ala Ile Met
Ala Pro Lys Leu Arg Thr Ala Phe 515 520 525Ala Ala Leu Ile Asn Asp
Arg Ser Glu Val Ala Leu Trp Ala Tyr Ser 530 535 540Gln Leu Ala Glu
Gln Gly Tyr Glu Thr Ala Gln Val Ser Ala Ala Tyr545 550 555 560Leu
Met Tyr Gln Leu Pro Tyr Glu Phe Glu Asp Pro Pro Arg Thr Thr 565 570
575Asp Gln Arg Lys Thr Leu Ala Ile Ser Tyr Tyr Thr Arg Ala Phe Lys
580 585 590Gln Gly Asn Ile Asp Ala Gly Val Val Ala Gly Asp Ile Tyr
Phe Gln 595 600 605Met Gln Asn Tyr Ser Lys Ala Met Ala Leu Tyr Gln
Gly Ala Ala Leu 610 615 620Lys Tyr Ser Ile Gln Ala Ile Trp Asn Leu
Gly Tyr Met His Glu His625 630 635 640Gly Leu Gly Val Asn Arg Asp
Phe His Leu Ala Lys Arg Tyr Tyr Asp 645 650 655Gln Val Ser Glu His
Asp His Arg Phe Tyr Leu Ala Ser Lys Leu Ser 660 665 670Val Leu Lys
Leu His Leu Lys Ser Trp Leu Thr Trp Ile Thr Arg Glu 675 680 685Lys
Val Asn Tyr Trp Lys Pro Ser Ser Pro Leu Asn Pro Asn Glu Asp 690 695
700Thr Gln His Ser Lys Thr Ser Trp Tyr Lys Gln Leu Thr Lys Ile
Leu705 710 715 720Gln Arg Met Arg His Lys Glu Asp Ser Asp Lys Ala
Ala Glu Asp Ser 725 730 735His Lys His Arg Thr Val Val Gln Asn Gly
Ala Asn His Arg Gly Asp 740 745 750Asp Gln Glu Glu Ala Ser Glu Ile
Leu Gly Phe Gln Met Glu Asp Leu 755 760 765Val Thr Met Gly Cys Ile
Leu Gly Ile Phe Leu Leu Ser Ile Leu Met 770 775 780Ser Thr Leu Ala
Ala Arg Arg Gly Trp Asn Val Arg Phe Asn Gly Ala785 790 795 800Gln
Leu Asn Ala Asn Gly Asn Arg Gln Gln Glu Gln Gln Gln Gln Gln 805 810
815Gln Ala Gln Gly Pro Pro Gly Trp Asp Phe Asn Val Gln Ile Phe Ala
820 825 830Ile 542502DNASaccharomyces cerevisiae 54atgataacac
tcttattata cctgtgcgta atatgtaacg caatagtgtt aataagggct 60gattcgatag
cggacccttg gcctgaagcg cgacatctac taaataccat agctaagtcc
120agagacccaa tgaaagaagc tgctatggaa cccaatgcag atgaatttgt
tggattctat 180gtaccgatgg attattcccc acgtaatgag gaaaaaaact
accagagcat ttggcaaaac 240gaaatcacag attctcaacg tcatatttat
gaattacttg tacaatcaag tgaacaattc 300aacaactcag aagcaacata
tacacttagc cagattcacc tttggagtca atataatttc 360ccgcataata
tgactttggc acacaaatac ttagaaaaat tcaatgatct aacccacttc
420accaatcatt cggccatctt cgacttagct gtgatgtatg ccactggggg
atgtgcttct 480ggtaatgatc aaaccgtgat ccctcaggat tctgctaaag
cactgctata ttaccaaagg 540gctgcccaac tagggaattt aaaggctaag
caagtgctag cttataaata ctattctggc 600ttcaatgtcc cacgaaattt
tcataaatct ttagtattgt acagggacat tgctgaacag 660ctgagaaagt
cgtactccag ggacgaatgg gatattgtct tcccctattg ggaaagttac
720aacgtgagaa tatcggattt tgagagtggc ctattaggta aaggtttgaa
ttccgttcca 780tcttctacag taaggaaaag aactacgaga ccagatattg
gttcaccctt tattgcgcaa 840gttaacggtg tacagatgac cttgcaaatc
gaaccgatgg gtaggttcgc tttcaacggt 900aacgatggca acataaatgg
cgacgaagat gacgaggatg ccagtgaaag acgaatcatt 960cggatatatt
atgcagcttt gaatgattat aaaggaacat attcacaaag cagaaattgt
1020gagcgcgcca aaaacttgtt ggaattaacg tacaaggaat ttcagcctca
tgtcgacaat 1080ttggatcctt tgcaagtatt ttactacgtc cgttgcttac
aattattggg gcacatgtat 1140ttcaccggcg aaggctcctc gaagcctaat
attcatatgg ccgaagagat cctgaccacg 1200tcgctagaaa taagcagaag
ggcacaggga cctataggta gagcgtgcat agatctgggc 1260ttaataaatc
aatacatcac aaacaatatt tctcaagcaa tttcgtatta tatgaaagct
1320atgaaaacac aagctaacaa tggaatcgta gaattccaat tatccaaatt
ggccacttca 1380ttccctgaag aaaaaatcgg cgacccattt aacttaatgg
aaactgccta cttgaatgga 1440ttcattccag ccatatatga gtttgcagta
atgatcgaat ctggaatgaa cagtaagagt 1500agtgtggaaa acactgctta
cctgttcaaa acattcgttg acaaaaacga agctattatg 1560gcacctaaac
tgaggacagc atttgccgca ttaatcaacg atcgttcaga agtggcttta
1620tgggcttatt cccaactagc cgagcaaggc tacgagactg ctcaagtctc
tgccgcctac 1680ttaatgtacc agttgccata tgagtttgag gatcctccaa
gaaccacaga tcagagaaaa 1740actttggcaa tttcctacta tacaagagcg
tttaaacagg gaaatataga tgctggtgtt 1800gtcgcgggag atatctattt
tcagatgcag aattacagta aagctatggc tctttatcag 1860ggtgcagctt
tgaagtactc tatacaggct atctggaact tagggtacat gcatgagcat
1920gggctaggtg taaacagaga tttccatctt gctaaacgtt actacgacca
agtttcagaa 1980cacgatcata gattttactt ggcttccaaa ttgagtgttt
taaaattaca cctaaagtca 2040tggttgactt ggatcaccag agaaaaagta
aactactgga aaccttcctc gccacttaac 2100cctaacgaag atactcagca
ctcgaagact tcatggtaca agcaattgac gaagattcta 2160caaagaatga
gacataagga ggatagtgac aaagctgcgg aagattctca caaacacaga
2220actgtagtgc agaatggagc taaccatagg ggtgacgacc aagaggaggc
ttccgagatt 2280ttgggcttcc aaatggagga tcttgttacg atgggatgta
tcttggggat attcctatta 2340agtatattaa tgagtacact ggcggcccgt
agaggctgga atgtccgttt caatggagca 2400caattaaatg caaatggtaa
ccggcagcaa gagcaacaac aacaacaaca agcacaaggt 2460cccccgggct
gggacttcaa tgttcagata ttcgccatat ga 250255165PRTSaccharomyces
cerevisiae 55Met Ser Lys Thr Ala Gln Lys Arg Leu Leu Lys Glu Leu
Gln Gln Leu1 5 10 15Ile Lys Asp Ser Pro Pro Gly Ile Val Ala Gly Pro
Lys Ser Glu Asn 20 25 30Asn Ile Phe Ile Trp Asp Cys Leu Ile Gln Gly
Pro Pro Asp Thr Pro 35 40 45Tyr Ala Asp Gly Val Phe Asn Ala Lys Leu
Glu Phe Pro Lys Asp Tyr 50 55 60Pro Leu Ser Pro Pro Lys Leu Thr Phe
Thr Pro Ser Ile Leu His Pro65 70 75 80Asn Ile Tyr Pro Asn Gly Glu
Val Cys Ile Ser Ile Leu His Ser Pro 85 90 95Gly Asp Asp Pro Asn Met
Tyr Glu Leu Ala Glu Glu Arg Trp Ser Pro 100 105 110Val Gln Ser Val
Glu Lys Ile Leu Leu Ser Val Met Ser Met Leu Ser 115 120 125Glu Pro
Asn Ile Glu Ser Gly Ala Asn Ile Asp Ala Cys Ile Leu Trp 130 135
140Arg Asp Asn Arg Pro Glu Phe Glu Arg Gln Val Lys Leu Ser Ile
Leu145 150 155 160Lys Ser Leu Gly Phe 16556498DNASaccharomyces
cerevisiae 56atgtcgaaaa ccgctcagaa acgtctcctc aaggagcttc aacagttaat
taaagattct 60ccacctggta tagtggctgg tcccaaatcg gagaataaca tattcatttg
ggactgccta 120attcaagggc ctccagatac gccatacgct gatggtgttt
ttaatgctaa gctagagttt 180cctaaagact atccgttatc tccacctaaa
cttactttca cacccagcat actacatcca 240aatatttatc caaatgggga
agtgtgcata tccattctac actcccctgg tgatgatcct 300aacatgtacg
aattagcgga agaaagatgg tcgccagtgc aaagtgtaga aaaaattcta
360ttaagtgtta tgagcatgtt
gagtgagccc aatatcgaaa gtggtgccaa cattgatgct 420tgcatcttgt
ggagagataa tagacctgaa tttgagagac aggtaaagtt atccattttg
480aaatcattag gattctga 49857926PRTSaccharomyces cerevisiae 57Met
Glu Gln Asn Ile Ile Ser Thr Ile Arg Asp Glu Cys Ile Arg His1 5 10
15Arg Ser Lys Tyr Leu Thr Ile Ala Gln Leu Thr Ala Ile Ala Glu Ala
20 25 30Lys Ile Asn Glu Phe Ile Ile Thr Gly Lys Ala Lys Asp Gln Asp
Leu 35 40 45Ser Ser Leu Leu Asp Lys Cys Ile Asp Ile Leu Ser Ile Tyr
Lys Lys 50 55 60Asn Ser Lys Asp Ile Lys Asn Ile Ile Ser Cys Lys Asn
Lys Gly Ala65 70 75 80Met Ile Ser Ser Asn Ser Val Met Ile Ile Gln
Leu Asn Tyr Val Tyr 85 90 95Tyr Lys Val Ile His Ile Ile Val Thr Thr
Asn Ile Pro His Leu Ser 100 105 110Glu Phe Ala Lys Ile Lys Leu His
Lys Ser Thr Ser Asp Glu Gly Asn 115 120 125Gly Asn Asn Asn Asn Asn
Glu Phe Gln Leu Met Asn Ile Tyr Asn Thr 130 135 140Leu Leu Glu Thr
Leu Leu Lys Asp Glu Asn Ile Ala Lys Ile Lys Ser145 150 155 160Phe
Ile Lys Ser Ser Ile Lys Gln Thr Lys Leu Asn His Glu Gln Glu 165 170
175Glu Cys Asn Leu Met Arg Thr Gly Ser Tyr Ile Thr Ser Asn Gln Leu
180 185 190Asn Ser Leu Ile Ser Ser Ser Ala Asn Ser Ala Ser Ser Gln
Met Glu 195 200 205Ile Leu Leu Ile Asp Ile Arg Ser Arg Leu Glu Phe
Asn Lys Ser His 210 215 220Ile Asp Thr Lys Asn Ile Ile Cys Leu Glu
Pro Ile Ser Phe Lys Met225 230 235 240Ser Tyr Ser Asp His Asp Leu
Glu Lys Lys Ser Leu Ile Thr Ser Pro 245 250 255Asn Ser Glu Ile Lys
Met Phe Gln Ser Arg Asn Leu Phe Lys Phe Ile 260 265 270Ile Leu Tyr
Thr Asp Ala Asn Glu Tyr Asn Val Lys Gln Gln Ser Val 275 280 285Leu
Leu Asp Ile Leu Val Asn His Ser Phe Glu Lys Pro Ile Ser Asp 290 295
300Asp Phe Thr Lys Ile Phe Ile Leu Glu Ser Gly Phe Pro Gly Trp
Leu305 310 315 320Lys Ser Asn Tyr Gly Arg Gln Val Ser Ser Ser Phe
Pro Ser Asn Asn 325 330 335Asn Ile Lys Asp Asp Ser Val Tyr Ile Asn
Gly Asn Thr Ser Gly Leu 340 345 350Ser Leu Gln His Leu Pro Lys Met
Ser Pro Ser Ile Arg His Ser Met 355 360 365Asp Asp Ser Met Lys Glu
Met Leu Val Ala Pro Thr Pro Leu Asn His 370 375 380Leu Gln Gln Gln
Gln Gln Gln Gln Ser Asp Asn Asp His Val Leu Lys385 390 395 400Arg
Ser Ser Ser Phe Lys Lys Leu Phe Ser Asn Tyr Thr Ser Pro Asn 405 410
415Pro Lys Asn Ser Asn Ser Asn Leu Tyr Ser Ile Ser Ser Leu Ser Ile
420 425 430Ser Ser Ser Pro Ser Pro Leu Pro Leu His Ser Pro Asp Pro
Val Lys 435 440 445Gly Asn Ser Leu Pro Ile Asn Tyr Pro Glu Thr Pro
His Leu Trp Lys 450 455 460Asn Ser Glu Thr Asp Phe Met Thr Asn Gln
Arg Glu Gln Leu Asn His465 470 475 480Asn Ser Phe Ala His Ile Ala
Pro Ile Asn Thr Lys Ala Ile Thr Ser 485 490 495Pro Ser Arg Thr Ala
Thr Pro Lys Leu Gln Arg Phe Pro Gln Thr Ile 500 505 510Ser Met Asn
Leu Asn Met Asn Ser Asn Gly His Ser Ser Ala Thr Ser 515 520 525Thr
Ile Gln Pro Ser Cys Leu Ser Leu Ser Asn Asn Asp Ser Leu Asp 530 535
540His Thr Asp Val Thr Pro Thr Ser Ser His Asn Tyr Asp Leu Asp
Phe545 550 555 560Ala Val Gly Leu Glu Asn Leu Gly Asn Ser Cys Tyr
Met Asn Cys Ile 565 570 575Ile Gln Cys Ile Leu Gly Thr His Glu Leu
Thr Gln Ile Phe Leu Asp 580 585 590Asp Ser Tyr Ala Lys His Ile Asn
Ile Asn Ser Lys Leu Gly Ser Lys 595 600 605Gly Ile Leu Ala Lys Tyr
Phe Ala Arg Leu Val His Met Met Tyr Lys 610 615 620Glu Gln Val Asp
Gly Ser Lys Lys Ile Ser Ile Ser Pro Ile Lys Phe625 630 635 640Lys
Leu Ala Cys Gly Ser Val Asn Ser Leu Phe Lys Thr Ala Ser Gln 645 650
655Gln Asp Cys Gln Glu Phe Cys Gln Phe Leu Leu Asp Gly Leu His Glu
660 665 670Asp Leu Asn Gln Cys Gly Ser Asn Pro Pro Leu Lys Glu Leu
Ser Gln 675 680 685Glu Ala Glu Ala Arg Arg Glu Lys Leu Ser Leu Arg
Ile Ala Ser Ser 690 695 700Ile Glu Trp Glu Arg Phe Leu Thr Thr Asp
Phe Ser Val Ile Val Asp705 710 715 720Leu Phe Gln Gly Gln Tyr Ala
Ser Arg Leu Lys Cys Lys Val Cys Ser 725 730 735His Thr Ser Thr Thr
Tyr Gln Pro Phe Thr Val Leu Ser Ile Pro Ile 740 745 750Pro Lys Lys
Asn Ser Arg Asn Asn Ile Thr Ile Glu Asp Cys Phe Arg 755 760 765Glu
Phe Thr Lys Cys Glu Asn Leu Glu Val Asp Glu Gln Trp Leu Cys 770 775
780Pro His Cys Glu Lys Arg Gln Pro Ser Thr Lys Gln Leu Thr Ile
Thr785 790 795 800Arg Leu Pro Arg Asn Leu Ile Val His Leu Lys Arg
Phe Asp Asn Leu 805 810 815Leu Asn Lys Asn Asn Asp Phe Val Ile Tyr
Pro Phe Leu Leu Asp Leu 820 825 830Thr Pro Phe Trp Ala Asn Asp Phe
Asp Gly Val Phe Pro Pro Gly Val 835 840 845Asn Asp Asp Glu Leu Pro
Ile Arg Gly Gln Ile Pro Pro Phe Lys Tyr 850 855 860Glu Leu Tyr Gly
Val Ala Cys His Phe Gly Thr Leu Tyr Gly Gly His865 870 875 880Tyr
Thr Ala Tyr Val Lys Lys Gly Leu Lys Lys Gly Trp Leu Tyr Phe 885 890
895Asp Asp Thr Lys Tyr Lys Pro Val Lys Asn Lys Ala Asp Ala Ile Asn
900 905 910Ser Asn Ala Tyr Val Leu Phe Tyr His Arg Val Tyr Gly Val
915 920 925582781DNASaccharomyces cerevisiae 58atggagcaga
atattattag taccataagg gatgagtgta ttcgtcaccg gtcgaagtac 60cttacgatag
cacaactaac cgctattgca gaggctaaaa ttaacgaatt catcataact
120ggtaaggcaa aagatcaaga tttgagcagt cttctagata aatgcatcga
tattttatct 180atttacaaga agaactcgaa agatatcaaa aatattatat
cgtgcaaaaa taagggtgca 240atgattagtt caaattccgt aatgattatt
caattaaatt atgtttacta caaggtaatt 300cacattattg taacaaccaa
tattcctcat ttaagtgaat tcgccaagat taaattacat 360aagagcacga
gtgatgaggg caacggtaat aacaacaata atgaatttca actcatgaac
420atttacaaca ctttgctgga aaccttatta aaagatgaaa acattgcaaa
aataaaaagt 480ttcattaagt cttccataaa acaaacaaaa ttgaaccatg
agcaagaaga atgtaacctg 540atgagaacgg gttcctatat cacttccaat
caattaaact ccctaataag ttcatcagca 600aattctgctt cctcccaaat
ggagatacta ctgatagata tacgatcaag gttggaattc 660aacaagtcac
atattgatac aaaaaatatt atatgcctgg agcctatttc ttttaaaatg
720tcatattcag atcatgattt ggagaaaaaa tcattaatta cttctcctaa
tagtgagatt 780aaaatgtttc aaagtagaaa tcttttcaag tttatcattc
tctatacaga cgcaaacgaa 840tacaatgtta aacagcagtc tgtcctgttg
gacattctgg tgaatcattc ctttgaaaaa 900ccaatatccg atgactttac
caaaattttc attctggaat ctggttttcc aggttggctt 960aagtcaaatt
atgggaggca agtatcatca tcttttccat caaataacaa tattaaagat
1020gatagtgttt atattaatgg taacacttct ggcctaagtt tacaacattt
acctaagatg 1080tctcccagta taagacattc aatggacgac tctatgaaag
aaatgctagt tgcgcctact 1140ccattaaatc atcttcaaca acagcaacaa
cagcaatcag acaatgatca tgtgctaaaa 1200agatcttcaa gtttcaaaaa
attattctca aattatacgt ctcctaatcc gaagaattca 1260aattcaaact
tatattctat atcttcgttg tccatatcta gttcaccatc gcctttacct
1320ctacattcgc ctgacccagt taagggcaat tcattgccaa tcaattatcc
ggaaacgcca 1380catctttgga aaaacagtga gacagatttt atgacaaatc
aaagagaaca gttgaatcac 1440aactcttttg ctcacatagc tcctatcaac
acgaaggcca tcacttctcc atcaagaact 1500gccacaccga agttacaacg
cttcccgcaa acaattagta tgaaccttaa tatgaactcc 1560aatggacaca
gttctgccac ctctaccatt caaccttcgt gtctatcctt gtctaataat
1620gactctttag atcatacaga tgttacacca acttcttctc ataattatga
ccttgatttc 1680gcggttggtt tggaaaatct aggaaattcg tgttacatga
actgtatcat tcagtgtatc 1740ttaggtacac acgaattaac ccaaatcttt
ttggacgatt catatgctaa acacatcaat 1800attaatagta agttgggatc
gaaaggtatt ctggcaaaat attttgcaag gttggttcat 1860atgatgtata
aggaacaggt tgatggttca aagaaaattt ccatatcacc gataaaattt
1920aaattggcat gtggatctgt taactcatta tttaagactg catcccaaca
ggactgccaa 1980gagttttgcc aattccttct agatggtctt catgaagact
tgaaccaatg cggttcaaac 2040ccacctttga aggagctttc tcaagaagct
gaggcgagaa gagaaaaact gtctttgcga 2100attgcctcgt caattgagtg
ggaacgattc ttgactactg atttcagtgt tattgtcgac 2160ttatttcagg
gacaatacgc ctcacgacta aaatgtaaag tctgtagtca tacctcgaca
2220acataccaac cttttacggt gctgtcaatc cctattccta aaaaaaattc
ccgaaataat 2280attaccattg aagattgttt cagagagttc accaaatgtg
agaacttgga agtggatgag 2340caatggttgt gcccacattg tgaaaaaagg
cagccctcca cgaaacaatt gacaataacg 2400agattaccga ggaatctgat
agtccattta aagagatttg ataatttatt aaacaaaaat 2460aatgacttcg
tcatataccc ttttttgttg gacttgactc cattttgggc caatgatttt
2520gacggggttt ttcctccagg tgttaatgac gatgaactac caataagggg
acaaatacca 2580ccttttaagt atgaattata tggtgtagca tgccactttg
gtactttgta tggtggtcat 2640tatacagcct atgtgaaaaa gggattaaag
aagggatggc tatattttga tgataccaaa 2700tataaacctg tcaaaaacaa
agccgatgca attaactcta atgcatacgt tttgttttat 2760caccgcgtct
acggtgtttg a 278159238PRTSaccharomyces cerevisiae 59Met Glu Met Thr
Asp Phe Glu Leu Thr Ser Asn Ser Gln Ser Asn Leu1 5 10 15Ala Ile Pro
Thr Asn Phe Lys Ser Thr Leu Pro Pro Arg Lys Arg Ala 20 25 30Lys Thr
Lys Glu Glu Lys Glu Gln Arg Arg Ile Glu Arg Ile Leu Arg 35 40 45Asn
Arg Arg Ala Ala His Gln Ser Arg Glu Lys Lys Arg Leu His Leu 50 55
60Gln Tyr Leu Glu Arg Lys Cys Ser Leu Leu Glu Asn Leu Leu Asn Ser65
70 75 80Val Asn Leu Glu Lys Leu Ala Asp His Glu Asp Ala Leu Thr Cys
Ser 85 90 95His Asp Ala Phe Val Ala Ser Leu Asp Glu Tyr Arg Asp Phe
Gln Ser 100 105 110Thr Arg Gly Ala Ser Leu Asp Thr Arg Ala Ser Ser
His Ser Ser Ser 115 120 125Asp Thr Phe Thr Pro Ser Pro Leu Asn Cys
Thr Met Glu Pro Ala Thr 130 135 140Leu Ser Pro Lys Ser Met Arg Asp
Ser Ala Ser Asp Gln Glu Thr Ser145 150 155 160Trp Glu Leu Gln Met
Phe Lys Thr Glu Asn Val Pro Glu Ser Thr Thr 165 170 175Leu Pro Ala
Val Asp Asn Asn Asn Leu Phe Asp Ala Val Ala Ser Pro 180 185 190Leu
Ala Asp Pro Leu Cys Asp Asp Ile Ala Gly Asn Ser Leu Pro Phe 195 200
205Asp Asn Ser Ile Asp Leu Asp Asn Trp Arg Asn Pro Glu Ala Gln Ser
210 215 220Gly Leu Asn Ser Phe Glu Leu Asn Asp Phe Phe Ile Thr
Ser225 230 23560717DNASaccharomyces cerevisiae 60atggaaatga
ctgattttga actaactagt aattcgcaat cgaacttggc tatccctacc 60aacttcaagt
cgactctgcc tccaaggaaa agagccaaga caaaagagga aaaggaacag
120cgaaggatcg agcgtatttt gagaaacaga agagctgctc accagagcag
agagaaaaaa 180agactacatc tgcagtatct cgagagaaaa tgttctcttt
tggaaaattt actgaacagc 240gtcaaccttg aaaaactggc tgaccacgaa
gacgcgttga cttgcagcca cgacgctttt 300gttgcttctc ttgacgagta
cagggatttc cagagcacga ggggcgcttc actggacacc 360agggccagtt
cgcactcgtc gtctgatacg ttcacacctt cacctctgaa ctgtacaatg
420gagcctgcga ctttgtcgcc caagagtatg cgcgattccg cgtcggacca
agagacttca 480tgggagctgc agatgtttaa gacggaaaat gtaccagagt
cgacgacgct acctgccgta 540gacaacaaca atttgtttga tgcggtggcc
tcgccgttgg cagacccact ctgcgacgat 600atagcgggaa acagtctacc
ctttgacaat tcaattgatc ttgacaattg gcgtaatcca 660gaagcgcagt
caggtttgaa ttcatttgaa ttgaatgatt tcttcatcac ttcatga
71761663PRTSaccharomyces cerevisiae 61Met Pro Thr Asn Tyr Glu Tyr
Asp Glu Ala Ser Glu Thr Trp Pro Ser1 5 10 15Phe Ile Leu Thr Gly Leu
Leu Met Val Val Gly Pro Met Thr Leu Leu 20 25 30Gln Ile Tyr Gln Ile
Phe Phe Gly Ala Asn Ala Glu Asp Gly Asn Ser 35 40 45Gly Lys Ser Lys
Glu Phe Asn Glu Glu Val Phe Lys Asn Leu Asn Glu 50 55 60Glu Tyr Thr
Ser Asp Glu Ile Lys Gln Phe Arg Arg Lys Phe Asp Lys65 70 75 80Asn
Ser Asn Lys Lys Ser Lys Ile Trp Ser Arg Arg Asn Ile Ile Ile 85 90
95Ile Val Gly Trp Ile Leu Val Ala Ile Leu Leu Gln Arg Ile Asn Ser
100 105 110Asn Asp Ala Ile Lys Asp Ala Ala Thr Lys Leu Phe Asp Pro
Tyr Glu 115 120 125Ile Leu Gly Ile Ser Thr Ser Ala Ser Asp Arg Asp
Ile Lys Ser Ala 130 135 140Tyr Arg Lys Leu Ser Val Lys Phe His Pro
Asp Lys Leu Ala Lys Gly145 150 155 160Leu Thr Pro Asp Glu Lys Ser
Val Met Glu Glu Thr Tyr Val Gln Ile 165 170 175Thr Lys Ala Tyr Glu
Ser Leu Thr Asp Glu Leu Val Arg Gln Asn Tyr 180 185 190Leu Lys Tyr
Gly His Pro Asp Gly Pro Gln Ser Thr Ser His Gly Ile 195 200 205Ala
Leu Pro Arg Phe Leu Val Asp Gly Ser Ala Ser Pro Leu Leu Val 210 215
220Val Cys Tyr Val Ala Leu Leu Gly Leu Ile Leu Pro Tyr Phe Val
Ser225 230 235 240Arg Trp Trp Ala Arg Thr Gln Ser Tyr Thr Lys Lys
Gly Ile His Asn 245 250 255Val Thr Ala Ser Asn Phe Val Ser Asn Leu
Val Asn Tyr Lys Pro Ser 260 265 270Glu Ile Val Thr Thr Asp Leu Ile
Leu His Trp Leu Ser Phe Ala His 275 280 285Glu Phe Lys Gln Phe Phe
Pro Asp Leu Gln Pro Thr Asp Phe Glu Lys 290 295 300Leu Leu Gln Asp
His Ile Asn Arg Arg Asp Ser Gly Lys Leu Asn Asn305 310 315 320Ala
Lys Phe Arg Ile Val Ala Lys Cys His Ser Leu Leu His Gly Leu 325 330
335Leu Asp Ile Ala Cys Gly Phe Arg Asn Leu Asp Ile Ala Leu Gly Ala
340 345 350Ile Asn Thr Phe Lys Cys Ile Val Gln Ala Val Pro Leu Thr
Pro Asn 355 360 365Cys Gln Ile Leu Gln Leu Pro Asn Val Asp Lys Glu
His Phe Ile Thr 370 375 380Lys Thr Gly Asp Ile His Thr Leu Gly Lys
Leu Phe Thr Leu Glu Asp385 390 395 400Ala Lys Ile Gly Glu Val Leu
Gly Ile Lys Asp Gln Ala Lys Leu Asn 405 410 415Glu Thr Leu Arg Val
Ala Ser His Ile Pro Asn Leu Lys Ile Ile Lys 420 425 430Ala Asp Phe
Leu Val Pro Gly Glu Asn Gln Val Thr Pro Ser Ser Thr 435 440 445Pro
Tyr Ile Ser Leu Lys Val Leu Val Arg Ser Ala Lys Gln Pro Leu 450 455
460Ile Pro Thr Ser Leu Ile Pro Glu Glu Asn Leu Thr Glu Pro Gln
Asp465 470 475 480Phe Glu Ser Gln Arg Asp Pro Phe Ala Met Met Ser
Lys Gln Pro Leu 485 490 495Val Pro Tyr Ser Phe Ala Pro Phe Phe Pro
Thr Lys Arg Arg Gly Ser 500 505 510Trp Cys Cys Leu Val Ser Ser Gln
Lys Asp Gly Lys Ile Leu Gln Thr 515 520 525Pro Ile Ile Ile Glu Lys
Leu Ser Tyr Lys Asn Leu Asn Asp Asp Lys 530 535 540Asp Phe Phe Asp
Lys Arg Ile Lys Met Asp Leu Thr Lys His Glu Lys545 550 555 560Phe
Asp Ile Asn Asp Trp Glu Ile Gly Thr Ile Lys Ile Pro Leu Gly 565 570
575Gln Pro Ala Pro Glu Thr Val Gly Asp Phe Phe Phe Arg Val Ile Val
580 585 590Lys Ser Thr Asp Tyr Phe Thr Thr Asp Leu Asp Ile Thr Met
Asn Met 595 600 605Lys Val Arg Asp Ser Pro Ala Val Glu Gln Val Glu
Val Tyr Ser Glu 610 615 620Glu Asp Asp Glu Tyr Ser Thr Asp Asp Asp
Glu Thr Glu Ser Asp Asp625 630 635 640Glu Ser Asp Ala Ser Asp Tyr
Thr Asp Ile Asp Thr Asp Thr Glu Ala 645 650 655Glu Asp Asp Glu Ser
Pro Glu 660621992DNASaccharomyces cerevisiae 62atgcctacaa
attacgagta tgatgaggct agtgagacgt ggccgtcctt cattttaacg 60gggctcttga
tggtcgtcgg
gcctatgaca ctgcttcaaa tataccaaat tttttttggg 120gccaatgctg
aagatgggaa ttcagggaag agtaaggagt ttaatgagga agttttcaag
180aacttgaatg aagaatacac cagtgatgaa atcaaacaat ttagaaggaa
gtttgataaa 240aatagtaata agaagtccaa aatatggagc aggagaaata
ttataattat tgtgggttgg 300atcttagttg caattcttct gcaaaggatt
aatagtaatg acgcgattaa agacgctgct 360acaaaattat ttgatcctta
tgaaatcctt ggtatctcta ctagtgcttc cgatagagac 420atcaaatctg
cttatagaaa attatctgtt aaatttcatc cagataaatt agcaaagggc
480ctaacacctg atgagaaaag tgtgatggaa gaaacttatg ttcagattac
gaaggcttac 540gaatccctta ctgacgaatt ggttaggcaa aactatttga
aatacggtca tccagatggc 600ccacaatcta cttcacatgg tatcgctcta
ccaagatttt tggtagatgg aagtgcatct 660ccattattag tggtttgtta
tgttgcgcta ctaggtttaa tcttgccata ttttgttagt 720agatggtggg
caagaacaca atcgtatact aagaagggaa tacataatgt gacggcttct
780aattttgtta gtaacttagt caattacaag ccatctgaga ttgtcaccac
agatttgatc 840ttacactggt tatcatttgc tcatgaattt aaacaattct
tcccggattt gcaaccaacg 900gattttgaaa aacttttgca agatcatatt
aaccgcagag atagtggtaa acttaacaat 960gcgaaattta gaatagtggc
caaatgtcac tctttgttac acggtttatt ggatattgct 1020tgtggattca
gaaatttaga tattgcattg ggtgcaatca atactttcaa gtgtattgtt
1080caggctgtac cattaacacc aaactgtcaa atccttcaat tgccgaacgt
agataaagag 1140cactttatta ccaaaaccgg agatattcat acattaggta
aattgtttac tttagaagat 1200gccaagattg gtgaggttct tggaataaag
gatcaagcaa agttaaacga aactttgaga 1260gttgcatcgc atattccaaa
tctaaagatc atcaaggcag acttccttgt cccaggtgag 1320aaccaagtaa
caccatcatc taccccatac atttctttga aagtactggt tcgttctgct
1380aaacagccat tgataccaac tagcttaatt cctgaagaaa atttaacaga
acctcaagat 1440tttgaatctc aaagagatcc atttgctatg atgagtaaac
agccactcgt cccatattcc 1500tttgcaccat ttttccctac aaagagacgt
gggagttggt gctgtctggt aagttctcaa 1560aaagatggta aaatacttca
aacgccaatt atcattgaaa agctatctta caagaacttg 1620aacgatgaca
aagatttctt tgataagagg ataaaaatgg atttaaccaa acacgaaaaa
1680ttcgatataa atgattggga aatcgggacc ataaaaattc cattaggtca
gcctgcacct 1740gaaactgttg gtgatttctt ttttagagta atcgttaaat
ccacagatta tttcactaca 1800gatttggata ttaccatgaa tatgaaagtt
cgtgattctc ctgcagtgga acaagtagag 1860gtgtattctg aggaggatga
tgagtactct actgatgacg acgaaaccga aagtgatgat 1920gaaagtgatg
ctagcgatta tactgatatc gatacggata cagaagctga agatgatgaa
1980tcaccagaat ag 199263409PRTSaccharomyces cerevisiae 63Met Val
Lys Glu Thr Lys Phe Tyr Asp Ile Leu Gly Val Pro Val Thr1 5 10 15Ala
Thr Asp Val Glu Ile Lys Lys Ala Tyr Arg Lys Cys Ala Leu Lys 20 25
30Tyr His Pro Asp Lys Asn Pro Ser Glu Glu Ala Ala Glu Lys Phe Lys
35 40 45Glu Ala Ser Ala Ala Tyr Glu Ile Leu Ser Asp Pro Glu Lys Arg
Asp 50 55 60Ile Tyr Asp Gln Phe Gly Glu Asp Gly Leu Ser Gly Ala Gly
Gly Ala65 70 75 80Gly Gly Phe Pro Gly Gly Gly Phe Gly Phe Gly Asp
Asp Ile Phe Ser 85 90 95Gln Phe Phe Gly Ala Gly Gly Ala Gln Arg Pro
Arg Gly Pro Gln Arg 100 105 110Gly Lys Asp Ile Lys His Glu Ile Ser
Ala Ser Leu Glu Glu Leu Tyr 115 120 125Lys Gly Arg Thr Ala Lys Leu
Ala Leu Asn Lys Gln Ile Leu Cys Lys 130 135 140Glu Cys Glu Gly Arg
Gly Gly Lys Lys Gly Ala Val Lys Lys Cys Thr145 150 155 160Ser Cys
Asn Gly Gln Gly Ile Lys Phe Val Thr Arg Gln Met Gly Pro 165 170
175Met Ile Gln Arg Phe Gln Thr Glu Cys Asp Val Cys His Gly Thr Gly
180 185 190Asp Ile Ile Asp Pro Lys Asp Arg Cys Lys Ser Cys Asn Gly
Lys Lys 195 200 205Val Glu Asn Glu Arg Lys Ile Leu Glu Val His Val
Glu Pro Gly Met 210 215 220Lys Asp Gly Gln Arg Ile Val Phe Lys Gly
Glu Ala Asp Gln Ala Pro225 230 235 240Asp Val Ile Pro Gly Asp Val
Val Phe Ile Val Ser Glu Arg Pro His 245 250 255Lys Ser Phe Lys Arg
Asp Gly Asp Asp Leu Val Tyr Glu Ala Glu Ile 260 265 270Asp Leu Leu
Thr Ala Ile Ala Gly Gly Glu Phe Ala Leu Glu His Val 275 280 285Ser
Gly Asp Trp Leu Lys Val Gly Ile Val Pro Gly Glu Val Ile Ala 290 295
300Pro Gly Met Arg Lys Val Ile Glu Gly Lys Gly Met Pro Ile Pro
Lys305 310 315 320Tyr Gly Gly Tyr Gly Asn Leu Ile Ile Lys Phe Thr
Ile Lys Phe Pro 325 330 335Glu Asn His Phe Thr Ser Glu Glu Asn Leu
Lys Lys Leu Glu Glu Ile 340 345 350Leu Pro Pro Arg Ile Val Pro Ala
Ile Pro Lys Lys Ala Thr Val Asp 355 360 365Glu Cys Val Leu Ala Asp
Phe Asp Pro Ala Lys Tyr Asn Arg Thr Arg 370 375 380Ala Ser Arg Gly
Gly Ala Asn Tyr Asp Ser Asp Glu Glu Glu Gln Gly385 390 395 400Gly
Glu Gly Val Gln Cys Ala Ser Gln 405641230DNASaccharomyces
cerevisiae 64atggttaaag aaactaagtt ttacgatatt ctaggtgttc cagtaactgc
cactgatgtc 60gaaattaaga aagcttatag aaaatgcgcc ttaaaatacc atccagataa
gaatccaagt 120gaggaagctg cagaaaagtt caaagaagct tcagcagcct
atgaaatttt atcagatcct 180gaaaagagag atatatatga ccaatttggt
gaagatggtc taagtggtgc tggtggcgct 240ggcggattcc caggtggtgg
attcggtttt ggtgacgata tcttttccca attctttggt 300gctggtggcg
cacaaagacc aagaggtccc caaagaggta aagatatcaa gcatgaaatt
360tctgcctcac ttgaagaatt atataagggt aggacagcta agttagccct
taacaaacag 420atcctatgta aagaatgtga aggtcgtggt ggtaagaaag
gcgccgtcaa gaagtgtacc 480agctgtaatg gtcaaggtat taaatttgta
acaagacaaa tgggtccaat gatccaaaga 540ttccaaacag agtgtgatgt
ctgtcacggt actggtgata tcattgatcc taaggatcgt 600tgtaaatctt
gtaacggtaa gaaagttgaa aacgaaagga agatcctaga agtccatgtc
660gaaccaggta tgaaagatgg tcaaagaatc gttttcaaag gtgaagctga
ccaagcccca 720gatgtcattc caggtgatgt tgtcttcata gtttctgaga
gaccacacaa gagcttcaag 780agagatggtg atgatttagt atatgaggct
gaaattgatc tattgactgc tatcgctggt 840ggtgaatttg cattggaaca
tgtttctggt gattggttaa aggtcggtat tgttccaggt 900gaagttattg
ccccaggtat gcgtaaggtc atcgaaggta aaggtatgcc aattccaaaa
960tacggtggct atggtaattt aatcatcaaa tttactatca agttcccaga
aaaccatttc 1020acatcagaag aaaacttgaa gaagttagaa gaaattttgc
ctccaagaat tgtcccagcc 1080attccaaaga aagctactgt ggacgaatgt
gtactcgcag actttgaccc agccaaatac 1140aacagaacac gggcctccag
gggtggtgca aactatgatt ccgatgaaga agaacaaggt 1200ggcgaaggtg
ttcaatgtgc atctcaatga 123065459PRTSaccharomyces cerevisiae 65Met
Ser Gly Ser Asp Arg Gly Asp Arg Leu Tyr Asp Val Leu Gly Val1 5 10
15Thr Arg Asp Ala Thr Val Gln Glu Ile Lys Thr Ala Tyr Arg Lys Leu
20 25 30Ala Leu Lys His His Pro Asp Lys Tyr Val Asp Gln Asp Ser Lys
Glu 35 40 45Val Asn Glu Ile Lys Phe Lys Glu Ile Thr Ala Ala Tyr Glu
Ile Leu 50 55 60Ser Asp Pro Glu Lys Lys Ser His Tyr Asp Leu Tyr Gly
Asp Asp Asn65 70 75 80Gly Ala Ala Ser Ser Gly Gly Ala Asn Gly Phe
Gly Asp Glu Asp Phe 85 90 95Met Asn Phe Phe Asn Asn Phe Phe Asn Asn
Gly Ser His Asp Gly Asn 100 105 110Asn Phe Pro Gly Glu Tyr Asp Ala
Tyr Glu Glu Gly Asn Ser Thr Ser 115 120 125Ser Lys Asp Ile Asp Ile
Asp Ile Ser Leu Thr Leu Lys Asp Leu Tyr 130 135 140Met Gly Lys Lys
Leu Lys Phe Asp Leu Lys Arg Gln Val Ile Cys Ile145 150 155 160Lys
Cys His Gly Ser Gly Trp Lys Pro Lys Arg Lys Ile His Val Thr 165 170
175His Asp Val Glu Cys Glu Ser Cys Ala Gly Lys Gly Ser Lys Glu Arg
180 185 190Leu Lys Arg Phe Gly Pro Gly Leu Val Ala Ser Gln Trp Val
Val Cys 195 200 205Glu Lys Cys Asn Gly Lys Gly Lys Tyr Thr Lys Arg
Pro Lys Asn Pro 210 215 220Lys Asn Phe Cys Pro Asp Cys Ala Gly Leu
Gly Leu Leu Ser Lys Lys225 230 235 240Glu Ile Ile Thr Val Asn Val
Ala Pro Gly His His Phe Asn Asp Val 245 250 255Ile Thr Val Lys Gly
Met Ala Asp Glu Glu Ile Asp Lys Thr Thr Cys 260 265 270Gly Asp Leu
Lys Phe His Leu Thr Glu Lys Gln Glu Asn Leu Glu Gln 275 280 285Lys
Gln Ile Phe Leu Lys Asn Phe Asp Asp Gly Ala Gly Glu Asp Leu 290 295
300Tyr Thr Ser Ile Thr Ile Ser Leu Ser Glu Ala Leu Thr Gly Phe
Glu305 310 315 320Lys Phe Leu Thr Lys Thr Phe Asp Asp Arg Leu Leu
Thr Leu Ser Val 325 330 335Lys Pro Gly Arg Val Val Arg Pro Gly Asp
Thr Ile Lys Ile Ala Asn 340 345 350Glu Gly Trp Pro Ile Leu Asp Asn
Pro His Gly Arg Cys Gly Asp Leu 355 360 365Tyr Val Phe Val His Ile
Glu Phe Pro Pro Asp Asn Trp Phe Asn Glu 370 375 380Lys Ser Glu Leu
Leu Ala Ile Lys Thr Asn Leu Pro Ser Ser Ser Ser385 390 395 400Cys
Ala Ser His Ala Thr Val Asn Thr Glu Asp Asp Ser Asn Leu Thr 405 410
415Asn Asn Glu Thr Ile Ser Asn Phe Arg Ile Ile His Thr Asp Asp Leu
420 425 430Pro Glu Gly Ile Arg Pro Phe Lys Pro Glu Ala Gln Asp Ser
Ala His 435 440 445Gln Lys Ala Arg Ser Ser Tyr Cys Cys Ile Gln 450
455661380DNASaccharomyces cerevisiae 66atgagtggca gtgatagagg
agaccggtta tacgatgtgt tgggggtgac gagagatgcg 60accgtgcaag agattaaaac
tgcttacaga aagcttgccc tgaaacatca tccggacaag 120tatgtggatc
aagactcaaa ggaggtaaat gaaatcaaat tcaaagagat cactgccgct
180tacgagatct tgagcgatcc ggagaagaaa tcacattacg acttgtatgg
tgatgataat 240ggtgccgcta gcagcggtgg cgctaatggc tttggagatg
aagattttat gaacttcttt 300aacaatttct tcaataatgg aagtcacgat
ggaaataatt tccctggcga gtatgatgcg 360tacgaagagg gcaactctac
aagctctaag gatatcgata tcgatatatc tcttactttg 420aaggatttgt
acatgggcaa gaagctgaag tttgatttaa agagacaggt catctgtata
480aagtgccacg gttctggctg gaaaccaaag aggaaaattc acgttacaca
cgatgtggaa 540tgtgaatcat gcgctggaaa gggttcaaag gaacgtctga
agaggtttgg tcccggtttg 600gtagcttcgc aatgggtggt ctgtgagaaa
tgtaatggta aggggaagta cactaaaaga 660cccaagaatc caaagaactt
ttgccccgat tgcgcaggct tggggctcct gtcaaagaag 720gaaatcatca
cagtgaacgt ggctccggga caccacttta acgacgtaat tacagtcaag
780gggatggcgg acgaggaaat cgataagacc acatgtggtg atttaaagtt
ccatctcact 840gaaaaacaag aaaacttgga gcagaagcaa atctttttga
agaactttga cgacggcgcc 900ggggaagatt tgtatacaag cattaccata
tcgttaagcg aggccttgac gggatttgag 960aaatttttga caaaaacctt
cgacgacagg ttactaacat tgagcgttaa acctggcaga 1020gtagtaagac
ctggtgacac catcaaaatc gccaatgaag gttggcccat tttagataac
1080cctcatggcc ggtgcggcga tctgtatgtt ttcgttcata ttgaatttcc
accagataac 1140tggttcaatg aaaaatcaga actactagca ataaaaacga
atctgccgtc atcttcatct 1200tgtgcctcac atgcgactgt aaatactgaa
gatgacagca acctgactaa caacgaaact 1260atatcaaatt tccggatcat
tcacacggac gatcttccag aagggataag gccgttcaag 1320ccagaagcac
aggattcagc gcatcagaaa gcaagaagtt cgtactgctg tatccaatga
138067528PRTSaccharomyces cerevisiae 67Met Gln Gln Asn Thr Ser Leu
Tyr Asp Ser Leu Asn Val Thr Ala Ala1 5 10 15Ala Ser Thr Ser Glu Ile
Lys Lys Ala Tyr Arg Asn Ala Ala Leu Lys 20 25 30Tyr His Pro Asp Lys
Asn Asn His Thr Glu Glu Ser Lys Arg Lys Phe 35 40 45Gln Glu Ile Cys
Gln Ala Tyr Glu Ile Leu Lys Asp Asn Arg Leu Arg 50 55 60Ala Leu Tyr
Asp Gln Tyr Gly Thr Thr Asp Glu Val Leu Ile Gln Glu65 70 75 80Gln
Gln Ala Gln Ala Gln Arg Gln Gln Ala Gly Pro Phe Ser Ser Ser 85 90
95Ser Asn Phe Asp Thr Glu Ala Met Ser Phe Pro Asp Leu Ser Pro Gly
100 105 110Asp Leu Phe Ala Gln Phe Phe Asn Ser Ser Ala Thr Pro Ser
Ser Asn 115 120 125Gly Ser Lys Ser Ser Phe Asn Phe Ser Phe Asn Asn
Ser Ser Thr Pro 130 135 140Ser Phe Ser Phe Val Asn Gly Ser Gly Val
Asn Asn Leu Tyr Ser Ser145 150 155 160Ser Ala Lys Tyr Asn Ser Asn
Asp Glu Asp His His Leu Asp Arg Gly 165 170 175Pro Asp Ile Lys His
Asn Leu Lys Cys Thr Leu Lys Glu Leu Tyr Met 180 185 190Gly Lys Thr
Ala Lys Leu Gly Leu Asn Arg Thr Arg Ile Cys Ser Val 195 200 205Cys
Asp Gly His Gly Gly Leu Lys Lys Cys Thr Cys Lys Thr Cys Lys 210 215
220Gly Gln Gly Ile Gln Thr Gln Thr Arg Arg Met Gly Pro Leu Val
Gln225 230 235 240Ser Trp Ser Gln Thr Cys Ala Asp Cys Gly Gly Ala
Gly Val Phe Val 245 250 255Lys Asn Lys Asp Ile Cys Gln Gln Cys Gln
Gly Leu Gly Phe Ile Lys 260 265 270Glu Arg Lys Ile Leu Gln Val Thr
Val Gln Pro Gly Ser Cys His Asn 275 280 285Gln Leu Ile Val Leu Thr
Gly Glu Gly Asp Glu Val Ile Ser Thr Lys 290 295 300Gly Gly Gly His
Glu Lys Val Ile Pro Gly Asp Val Val Ile Thr Ile305 310 315 320Leu
Arg Leu Lys Asp Pro Asn Phe Gln Val Ile Asn Tyr Ser Asn Leu 325 330
335Ile Cys Lys Lys Cys Lys Ile Asp Phe Met Thr Ser Leu Cys Gly Gly
340 345 350Val Val Tyr Ile Glu Gly His Pro Ser Gly Lys Leu Ile Lys
Leu Asp 355 360 365Ile Ile Pro Gly Glu Ile Leu Lys Pro Gly Cys Phe
Lys Thr Val Glu 370 375 380Asp Met Gly Met Pro Lys Phe Ile Asn Gly
Val Arg Ser Gly Phe Gly385 390 395 400His Leu Tyr Val Lys Phe Asp
Val Thr Tyr Pro Glu Arg Leu Glu Pro 405 410 415Glu Asn Ala Lys Lys
Ile Gln Asn Ile Leu Ala Asn Asp Lys Tyr Ile 420 425 430Lys Ala Glu
Arg Ser Thr Met Glu Thr Ala Asp Ser Asp Cys Tyr Cys 435 440 445Asp
Leu Glu Lys Ser Tyr Asp Ser Val Glu Glu His Val Leu Ser Ser 450 455
460Phe Glu Ala Pro Asn Leu Asn Asn Glu Val Ile Glu Asp Asp Asp
Leu465 470 475 480Gly Asp Leu Ile Asn Glu Arg Asp Ser Arg Lys Arg
Asn Asn Arg Arg 485 490 495Phe Asp Glu Ser Asn Ile Asn Asn Asn Asn
Glu Thr Lys Arg Asn Lys 500 505 510Tyr Ser Ser Pro Val Ser Gly Phe
Tyr Asp His Asp Ile Asn Gly Tyr 515 520 525681587DNASaccharomyces
cerevisiae 68atgcaacaaa acacgtcttt atatgactct ttgaacgtta ctgccgctgc
atccacatct 60gagattaaga aagcttacag gaacgctgca ttaaaatatc atcctgataa
aaacaatcat 120acagaagaat ccaagcgaaa gtttcaagag atatgccagg
catacgaaat acttaaagac 180aatcgtttaa gagctttgta tgaccagtac
ggtaccacag atgaagtcct gattcaagag 240cagcaggcgc aggcgcaacg
ccaacaagcc gggccgttca gttcatcctc aaatttcgat 300acggaagcaa
tgtcattccc ggatctatct ccaggtgatc ttttcgcgca gttttttaat
360agttctgcta ccccctcttc taatggctcc aaaagcagtt ttaattttag
cttcaataat 420agctctacgc cgagcttctc ctttgttaat ggcagtggcg
tgaacaatct gtactcctcg 480tcagcaaaat acaactccaa cgatgaggac
catcatttgg atagaggccc tgatatcaaa 540cataatctaa agtgcacatt
gaaggaactc tacatgggta agactgcaaa gttgggtttg 600aataggacaa
ggatttgcag tgtttgtgat gggcacggtg gtctaaagaa atgcacttgt
660aaaacatgca aagggcaagg tattcaaacc caaactaggc gtatgggacc
tctagtacaa 720agttggtctc aaacttgtgc agattgcggg ggtgccgggg
tttttgtcaa aaataaagat 780atttgccaac agtgccaagg tcttggcttc
attaaggaga ggaagattct acaagtcacc 840gttcaaccgg gatcgtgtca
taaccaactt atagtactta cgggcgaagg tgacgaagtt 900attagtacta
agggaggcgg tcacgaaaag gtaatacctg gtgacgtcgt tatcaccatt
960ttacgtttaa aagatccgaa tttccaggtt atcaactact ccaatttgat
atgtaagaag 1020tgcaaaatcg acttcatgac cagtttatgt ggaggcgtag
tttatattga agggcaccct 1080agcggtaagt tgatcaaact tgatattata
cctggcgaga tactgaagcc tggttgtttc 1140aagactgttg aggacatggg
gatgcccaag tttatcaacg gtgttcggag cggtttcggt 1200catctatatg
tcaaattcga tgtgacgtat ccagagagac tggaacctga aaatgctaag
1260aaaatacaaa atattctggc taatgataaa tacattaaag cagaacgttc
caccatggaa 1320accgcagatt cagactgcta ttgcgatttg gagaagtcat
atgacagtgt ggaagagcat 1380gtgttaagta gctttgaggc ccctaattta
aacaatgaag ttattgaaga cgacgacctt 1440ggtgatttga ttaatgaaag
agattctcgg aaaaggaaca accgtcgatt cgacgaaagt 1500aatattaata
ataataatga aacgaaacga aataaatatt cttcaccggt aagcggtttt
1560tatgaccatg atattaatgg atattga 158769352PRTSaccharomyces
cerevisiae 69Met Val Lys Glu Thr Lys Leu Tyr Asp Leu Leu Gly Val
Ser Pro Ser1 5 10 15Ala Asn Glu Gln Glu Leu Lys Lys Gly Tyr Arg Lys
Ala Ala
Leu Lys 20 25 30Tyr His Pro Asp Lys Pro Thr Gly Asp Thr Glu Lys Phe
Lys Glu Ile 35 40 45Ser Glu Ala Phe Glu Ile Leu Asn Asp Pro Gln Lys
Arg Glu Ile Tyr 50 55 60Asp Gln Tyr Gly Leu Glu Ala Ala Arg Ser Gly
Gly Pro Ser Phe Gly65 70 75 80Pro Gly Gly Pro Gly Gly Ala Gly Gly
Ala Gly Gly Phe Pro Gly Gly 85 90 95Ala Gly Gly Phe Ser Gly Gly His
Ala Phe Ser Asn Glu Asp Ala Phe 100 105 110Asn Ile Phe Ser Gln Phe
Phe Gly Gly Ser Ser Pro Phe Gly Gly Ala 115 120 125Asp Asp Ser Gly
Phe Ser Phe Ser Ser Tyr Pro Ser Gly Gly Gly Ala 130 135 140Gly Met
Gly Gly Met Pro Gly Gly Met Gly Gly Met His Gly Gly Met145 150 155
160Gly Gly Met Pro Gly Gly Phe Arg Ser Ala Ser Ser Ser Pro Thr Tyr
165 170 175Pro Glu Glu Glu Thr Val Gln Val Asn Leu Pro Val Ser Leu
Glu Asp 180 185 190Leu Phe Val Gly Lys Lys Lys Ser Phe Lys Ile Gly
Arg Lys Gly Pro 195 200 205His Gly Ala Ser Glu Lys Thr Gln Ile Asp
Ile Gln Leu Lys Pro Gly 210 215 220Trp Lys Ala Gly Thr Lys Ile Thr
Tyr Lys Asn Gln Gly Asp Tyr Asn225 230 235 240Pro Gln Thr Gly Arg
Arg Lys Thr Leu Gln Phe Val Ile Gln Glu Lys 245 250 255Ser His Pro
Asn Phe Lys Arg Asp Gly Asp Asp Leu Ile Tyr Thr Leu 260 265 270Pro
Leu Ser Phe Lys Glu Ser Leu Leu Gly Phe Ser Lys Thr Ile Gln 275 280
285Thr Ile Asp Gly Arg Thr Leu Pro Leu Ser Arg Val Gln Pro Val Gln
290 295 300Pro Ser Gln Thr Ser Thr Tyr Pro Gly Gln Gly Met Pro Thr
Pro Lys305 310 315 320Asn Pro Ser Gln Arg Gly Asn Leu Ile Val Lys
Tyr Lys Val Asp Tyr 325 330 335Pro Ile Ser Leu Asn Asp Ala Gln Lys
Arg Ala Ile Asp Glu Asn Phe 340 345 350701059DNASaccharomyces
cerevisiae 70atggtcaagg agacaaaact ttatgattta cttggagtat ctccaagtgc
taatgagcaa 60gaactgaaaa agggttatag aaaagcagct ctaaaatatc atccagataa
gccaacaggt 120gacacagaaa agtttaagga gatatcagag gcctttgaaa
ttttaaatga tcctcaaaaa 180agggaaatat atgatcaata cggtctcgag
gctgctagat ctggtggtcc aagctttggt 240cctggtggtc ctggcggtgc
tggaggtgct ggaggcttcc ctggcggtgc gggcggattc 300tccggaggac
atgcgttcag taatgaggat gctttcaata ttttttcaca attctttggc
360ggcagttccc cattcggtgg tgctgatgac agtggcttca gtttctctag
ttatccatct 420ggcggcggtg ctggtatggg aggtatgcct ggaggaatgg
gaggaatgca tggcggcatg 480ggaggtatgc ctggcggctt tagatcagca
tcaagctctc ccacgtatcc agaggaagaa 540acagttcaag ttaatttacc
agttagtcta gaagatttgt ttgttggtaa aaagaagtca 600tttaaaattg
gaagaaaggg cccacatggg gcctctgaaa agacacaaat tgacattcaa
660ttaaaaccgg gttggaaagc tggtaccaaa ataacataca agaaccaggg
tgattacaat 720cctcaaacgg gccgtagaaa gactttgcag tttgtcatcc
aggaaaagag ccatccaaac 780tttaaaagag acggtgatga cctaatttac
actctgccac tatctttcaa ggaatcattg 840ttaggttttt caaaaactat
ccaaacaatt gatggcagaa ccttaccttt gtcgagagta 900cagcctgtcc
aaccctcaca aacttctact tatcctggtc aaggtatgcc aactccaaag
960aacccatctc agagaggtaa tttgattgta aaatataaag tggactatcc
aatatcacta 1020aacgacgctc aaaaacgtgc tatagatgaa aatttttaa
105971432PRTSaccharomyces cerevisiae 71Met Val Val Asp Thr Glu Tyr
Tyr Asp Leu Leu Gly Val Ser Thr Thr1 5 10 15Ala Ser Ser Ile Glu Ile
Lys Lys Ala Tyr Arg Lys Lys Ser Ile Gln 20 25 30Glu His Pro Asp Lys
Asn Pro Asn Asp Pro Thr Ala Thr Glu Arg Phe 35 40 45Gln Ala Ile Ser
Glu Ala Tyr Gln Val Leu Gly Asp Asp Asp Leu Arg 50 55 60Ala Lys Tyr
Asp Lys Tyr Gly Arg Lys Glu Ala Ile Pro Gln Gly Gly65 70 75 80Phe
Glu Asp Ala Ala Glu Gln Phe Ser Val Ile Phe Gly Gly Asp Ala 85 90
95Phe Ala Ser Tyr Ile Gly Glu Leu Met Leu Leu Lys Asn Leu Gln Lys
100 105 110Thr Glu Glu Leu Asn Ala Glu Asp Glu Ala Glu Lys Glu Lys
Glu Asn 115 120 125Val Glu Thr Met Glu Glu Ser Pro Ala Asp Gly Lys
Thr Asn Gly Thr 130 135 140Thr Asn Ala Val Asp Ala Ala Leu Gly Asn
Thr Asn Glu Lys Asp Asp145 150 155 160Lys Asn Lys Ala Arg Thr Thr
Ser Gly Asn Leu Thr Val His Asp Gly 165 170 175Asn Lys Lys Asn Glu
Gln Val Gly Ala Glu Ala Lys Lys Lys Lys Thr 180 185 190Lys Leu Glu
Gln Phe Glu Glu Glu Gln Glu Val Glu Lys Gln Lys Arg 195 200 205Val
Asp Gln Leu Ser Lys Thr Leu Ile Glu Arg Leu Ser Ile Leu Thr 210 215
220Glu Ser Val Tyr Asp Asp Ala Cys Lys Asp Ser Phe Lys Lys Lys
Phe225 230 235 240Glu Glu Glu Ala Asn Leu Leu Lys Met Glu Ser Phe
Gly Leu Asp Ile 245 250 255Leu His Thr Ile Gly Asp Val Tyr Tyr Glu
Lys Ala Glu Ile Phe Leu 260 265 270Ala Ser Gln Asn Leu Phe Gly Met
Gly Gly Ile Phe His Ser Met Lys 275 280 285Ala Lys Gly Gly Val Phe
Met Asp Thr Leu Arg Thr Val Ser Ala Ala 290 295 300Ile Asp Ala Gln
Asn Thr Met Lys Glu Leu Glu Lys Met Lys Glu Ala305 310 315 320Ser
Thr Asn Asn Glu Pro Leu Phe Asp Lys Asp Gly Asn Glu Gln Ile 325 330
335Lys Pro Thr Thr Glu Glu Leu Ala Gln Gln Glu Gln Leu Leu Met Gly
340 345 350Lys Val Leu Ser Ala Ala Trp His Gly Ser Lys Tyr Glu Ile
Thr Ser 355 360 365Thr Leu Arg Gly Val Cys Lys Lys Val Leu Glu Asp
Asp Ser Val Ser 370 375 380Lys Lys Thr Leu Ile Arg Arg Ala Glu Ala
Met Lys Leu Leu Gly Glu385 390 395 400Val Phe Lys Lys Thr Phe Arg
Thr Lys Val Glu Gln Glu Glu Ala Gln 405 410 415Ile Phe Glu Glu Leu
Val Ala Glu Ala Thr Lys Lys Lys Arg His Thr 420 425
430721299DNASaccharomyces cerevisiae 72atggttgttg atactgagta
ttacgatttg ttaggtgtgt ctaccactgc atcttccatt 60gaaataaaaa aggcctatag
aaagaaatct attcaagagc atcctgataa gaatcccaat 120gaccccacgg
ctaccgaaag gtttcaagca atatccgaag cttatcaagt tttaggtgac
180gatgatcttc gcgcaaagta tgataagtat ggaagaaaag aagctattcc
tcagggcggc 240tttgaagatg cagctgaaca gttctctgtc atctttggtg
gagatgcgtt tgcctcatat 300attggcgaac tgatgctatt aaagaaccta
cagaaaactg aggagctaaa tgctgaagac 360gaagctgaaa aggagaagga
gaatgtggaa acaatggaag aatcacctgc agacggtaag 420acgaatggca
ccactaacgc tgttgatgca gcattgggca atactaacga aaaagatgac
480aaaaataagg cgaggacaac ttctggtaat ttaactgtac acgatggaaa
caagaaaaat 540gagcaggtag gagcagaagc taagaagaag aagacaaaat
tagagcagtt tgaggaagaa 600caagaggtag aaaagcaaaa aagagtagac
caattaagca aaacattgat tgaaagatta 660tcgatattaa cagaaagtgt
ctatgatgat gcatgtaaag attcctttaa aaaaaagttc 720gaagaggaag
ccaatctttt aaagatggaa tcatttggtc tggacatatt acacacaata
780ggcgacgttt actacgaaaa agctgaaatt tttcttgcat cccagaacct
gttcggaatg 840ggtggtatat ttcattctat gaaggctaaa gggggagtat
ttatggatac actaagaact 900gtttcggcag ccatagacgc tcagaatact
atgaaggagc ttgaaaaaat gaaagaagct 960agcacgaata atgagccttt
gtttgacaaa gacggaaatg agcaaattaa gccaaccact 1020gaggaactgg
cgcagcaaga gcagctattg atgggcaaag tattgtcggc tgcttggcat
1080ggttctaaat atgaaataac atccacttta cgtggcgttt gtaaaaaagt
actagaagat 1140gactcggtaa gtaagaaaac gcttatcaga agagctgaag
caatgaaact attgggtgaa 1200gtctttaaga aaactttcag aaccaaagtc
gaacaagaag aggcacagat ctttgaagaa 1260cttgtagcag aagctacaaa
aaagaagaga catacatga 129973433PRTSaccharomyces cerevisiae 73Met Phe
Ser Leu Pro Thr Leu Thr Ser Asp Ile Thr Val Glu Val Asn1 5 10 15Ser
Ser Ala Thr Lys Thr Pro Phe Val Arg Arg Pro Val Glu Pro Val 20 25
30Gly Lys Phe Phe Leu Gln His Ala Gln Arg Thr Leu Arg Asn His Thr
35 40 45Trp Ser Glu Phe Glu Arg Ile Glu Ala Glu Lys Asn Val Lys Thr
Val 50 55 60Asp Glu Ser Asn Val Asp Pro Asp Glu Leu Leu Phe Asp Thr
Glu Leu65 70 75 80Ala Asp Glu Asp Leu Leu Thr His Asp Ala Arg Asp
Trp Lys Thr Ala 85 90 95Asp Leu Tyr Ala Ala Met Gly Leu Ser Lys Leu
Arg Phe Arg Ala Thr 100 105 110Glu Ser Gln Ile Ile Lys Ala His Arg
Lys Gln Val Val Lys Tyr His 115 120 125Pro Asp Lys Gln Ser Ala Ala
Gly Gly Ser Leu Asp Gln Asp Gly Phe 130 135 140Phe Lys Ile Ile Gln
Lys Ala Phe Glu Thr Leu Thr Asp Ser Asn Lys145 150 155 160Arg Ala
Gln Tyr Asp Ser Cys Asp Phe Val Ala Asp Val Pro Pro Pro 165 170
175Lys Lys Gly Thr Asp Tyr Asp Phe Tyr Glu Ala Trp Gly Pro Val Phe
180 185 190Glu Ala Glu Ala Arg Phe Ser Lys Lys Thr Pro Ile Pro Ser
Leu Gly 195 200 205Asn Lys Asp Ser Ser Lys Lys Glu Val Glu Gln Phe
Tyr Ala Phe Trp 210 215 220His Arg Phe Asp Ser Trp Arg Thr Phe Glu
Phe Leu Asp Glu Asp Val225 230 235 240Pro Asp Asp Ser Ser Asn Arg
Asp His Lys Arg Tyr Ile Glu Arg Lys 245 250 255Asn Lys Ala Ala Arg
Asp Lys Lys Lys Thr Ala Asp Asn Ala Arg Leu 260 265 270Val Lys Leu
Val Glu Arg Ala Val Ser Glu Asp Pro Arg Ile Lys Met 275 280 285Phe
Lys Glu Glu Glu Lys Lys Glu Lys Glu Arg Arg Lys Trp Glu Arg 290 295
300Glu Ala Gly Ala Arg Ala Glu Ala Glu Ala Lys Ala Lys Ala Glu
Ala305 310 315 320Glu Ala Lys Ala Lys Ala Glu Ser Glu Ala Lys Ala
Asn Ala Ser Ala 325 330 335Lys Ala Asp Lys Lys Lys Ala Lys Glu Ala
Ala Lys Ala Ala Lys Lys 340 345 350Lys Asn Lys Arg Ala Ile Arg Asn
Ser Ala Lys Glu Ala Asp Tyr Phe 355 360 365Gly Asp Ala Asp Lys Ala
Thr Thr Ile Asp Glu Gln Val Gly Leu Ile 370 375 380Val Asp Ser Leu
Asn Asp Glu Glu Leu Val Ser Thr Ala Asp Lys Ile385 390 395 400Lys
Ala Asn Ala Ala Gly Ala Lys Glu Val Leu Lys Glu Ser Ala Lys 405 410
415Thr Ile Val Asp Ser Gly Lys Leu Pro Ser Ser Leu Leu Ser Tyr Phe
420 425 430Val 741302DNASaccharomyces cerevisiae 74atgttttctt
tacctaccct aacctcagac atcactgttg aagtcaacag ttccgctacc 60aaaaccccat
tcgtccgtcg tccggtcgaa ccggttggta agttcttttt gcaacatgct
120caaagaactt tgagaaacca cacctggtct gaatttgaaa gaattgaagc
tgaaaagaac 180gtcaaaaccg ttgatgaatc caatgtcgac ccagatgagt
tgttattcga cactgaattg 240gccgatgaag atttactgac tcatgatgct
agagactgga aaactgccga tttgtatgct 300gctatgggtt tgtctaagtt
gcgtttcaga gctactgaaa gtcaaatcat caaggctcac 360agaaaacaag
ttgtcaagta ccatccagac aagcaatctg ctgctggtgg tagtttggac
420caagatggct ttttcaagat tattcaaaag gcctttgaaa ctttgactga
ttccaacaag 480agagctcagt acgactcatg tgattttgtt gccgatgttc
ctcctccaaa gaagggtacc 540gattatgact tttatgaagc ttggggcccc
gttttcgaag ctgaagctcg tttttctaag 600aagactccta ttccttctct
aggtaacaaa gattcttcca agaaggaagt tgaacaattc 660tatgctttct
ggcacagatt tgactcctgg agaacctttg agttcttgga cgaagatgtc
720ccagatgact cttctaacag agaccacaag cgttacattg aaagaaagaa
caaggccgca 780agagacaaga agaagactgc tgataacgct agattggtca
aacttgttga aagagctgtc 840agtgaagatc cccgtatcaa aatgttcaaa
gaagaagaga agaaggaaaa ggaaagaaga 900aaatgggaaa gagaagccgg
tgccagagct gaagctgaag ctaaggccaa ggccgaagct 960gaagcgaagg
ctaaagctga atctgaagcc aaggctaacg cctccgcaaa agctgacaaa
1020aagaaggcta aggaagctgc taaggccgcc aagaaaaaga acaagagagc
catccgtaac 1080tctgctaagg aagctgacta ctttggtgat gctgacaagg
ccaccacgat tgacgaacaa 1140gttggtttga tcgttgacag tttgaatgac
gaagagttag tgtccaccgc cgataagatc 1200aaggccaatg ctgctggtgc
caaggaagtt ttgaaggaat ctgcaaagac tattgtcgat 1260tctggcaaac
taccatccag cttgttgtcc tacttcgtgt ga 130275668PRTSaccharomyces
cerevisiae 75Met Ser Asp Pro Phe Ala His Leu Leu Thr Ser Leu Lys
Asn Lys Asp1 5 10 15Ser Ala Ser Ala Ser Lys Glu Thr Thr Pro Gln Ser
Ser Asn Ser Pro 20 25 30Ser Ile Thr Gly Ser Ala Val Ala Asp Val Ala
Arg Thr Asp Lys Ser 35 40 45Pro Asn Asp Ser Leu His Ser Ile Ser Ala
Pro Pro Leu Ile Pro Ser 50 55 60Pro Lys Val Asp Phe Ser Ala Pro Pro
Leu Val Pro Thr Asn Ser Thr65 70 75 80Thr Lys Ser Asn Thr Ala Asn
Asn Thr Pro Pro Ser Ala Leu Ala Asn 85 90 95Thr Asp Asp Asp Phe Asn
Gln Leu Phe Gly Met Gly Thr Val Thr Thr 100 105 110Thr Asp Thr Ile
Gln Lys Pro Asp Glu Asp Tyr Tyr Gly Ser Lys Glu 115 120 125Asp His
Leu Tyr Asn Gly Asp Asp Ala Leu Val Asp Glu Val Lys Asp 130 135
140Met Glu Ile Ala Arg Leu Met Ser Leu Gly Leu Ser Ile Glu Glu
Ala145 150 155 160Thr Glu Phe Tyr Glu Asn Asp Val Thr Tyr Glu Arg
Tyr Leu Glu Ile 165 170 175Leu Lys Ser Lys Gln Lys Glu Arg Asn Asp
Leu Ala Ile Arg Lys Lys 180 185 190Glu Ser Gly Ile Lys Met Glu Lys
Ser Gly Leu Ser Asn Ile Val Gly 195 200 205Thr Asp Ser Asn Asn Leu
Phe Ser Met Ala Thr Asp Phe Phe Asn Lys 210 215 220Gly Lys Lys Leu
Val Asp Gln Trp Thr Ser Phe Pro Pro Glu Ala Asn225 230 235 240Asp
Arg Leu Asn Asn Tyr Ser Lys Thr His Asp Lys Val Glu Asp Tyr 245 250
255Asp Leu Pro Gln Val Asn Asp Ser Pro Asn Arg Ile Leu Phe Glu Asp
260 265 270Asn Glu Val Val Glu Asn Leu Pro Pro Ala Asp Asn Pro Asp
Gln Asp 275 280 285Leu Leu Thr Asp Phe Glu Thr Lys Ile Asp Ile Thr
Lys Arg Thr Ala 290 295 300Pro Asp Val Ser His Ser Ser Ser Pro Thr
Ser Gly Ile Leu Ile Glu305 310 315 320Glu Asn Ser Arg Arg Asn Glu
Pro Leu Ile Glu Asp Ser Leu Leu Asp 325 330 335Phe Ser Glu Gly Asn
Leu Thr Asn Ser Lys Ser Asn Glu Asp Ser Thr 340 345 350Leu Phe Asn
Glu Asn Ser Asn Thr Asp Ser Thr Ile Pro Ile Ser Asp 355 360 365Ile
Glu Leu Ser Gly Tyr Asn Glu Phe Lys Ala Lys Gly Thr Ser Leu 370 375
380Phe Lys Asn Gly Asp Tyr Ile Asn Ser Leu Gln Glu Tyr Glu Lys
Ser385 390 395 400Leu Asn Thr Leu Pro Leu Asn His Pro Leu Arg Ile
Ile Ala Leu Ser 405 410 415Asn Ile Ile Ala Ser Gln Leu Lys Ile Gly
Glu Tyr Ser Lys Ser Ile 420 425 430Glu Asn Ser Ser Met Ala Leu Glu
Leu Phe Pro Ser Ser Lys Ala Lys 435 440 445Trp Lys Asn Lys Ile Ser
Asn Ser Asp Pro Glu Arg Ser Phe Asn Asp 450 455 460Ile Trp Pro Lys
Ile Met Ile Arg Arg Ala Glu Ser Phe Glu His Leu465 470 475 480Glu
Ser Phe Lys Lys Ala Leu Glu Thr Tyr Gln Glu Leu Ile Lys Lys 485 490
495Asn Phe Phe Asp Asp Lys Ile Met Gln Gly Lys Arg Arg Cys Gln Asp
500 505 510Phe Ile Asn Pro Pro Pro Val Lys Lys Ser Met Pro Val Lys
Lys Lys 515 520 525Thr Thr Thr Thr Ser Pro Ala Thr Lys Lys Gln Asn
Leu Thr Ala Ser 530 535 540Ser Ser Asn Ser Pro Ile Ser Val Asp Ser
Thr Ser Glu Ile Lys Lys545 550 555 560Arg Glu Leu Glu Asn Ala Lys
Leu Ala Leu Tyr Asp Lys Val Phe Glu 565 570 575Lys Ile Ser Ser Trp
Lys Asp Gly Lys Asp Asp Asp Ile Arg His Leu 580 585 590Leu Ala Asn
Leu Ser Ser Leu Leu Thr Trp Cys Asn Trp Lys Asp Val 595 600 605Ser
Met Gln Asp Leu Val Met Pro Lys Arg Val Lys Ile Thr Tyr Met 610 615
620Lys Ala Val Ala Lys Thr His Pro Asp Lys Ile Pro Glu Ser Leu
Ser625 630 635 640Leu Glu
Asn Lys Met Ile Ala Glu Asn Ile Phe Ser Thr Leu Ser Ile 645 650
655Ala Trp Asp Lys Phe Lys Leu Gln Asn Asp Ile Asn 660
665762007DNASaccharomyces cerevisiae 76atgtcagatc catttgcaca
tttactgact tctttgaaga ataaggactc tgcatctgca 60tccaaggaaa caactcctca
gagcagcaat tcgccttcca ttactggttc cgctgttgca 120gatgttgcaa
ggacggataa aagccccaat gatagtctgc attcaatttc agctcctccg
180ctgataccgt caccgaaggt agatttttct gcacctcctt tggtcccaac
taatagcacc 240actaaatcta atactgccaa caacacacct ccctcggctc
ttgccaatac cgatgacgac 300ttcaatcaac tatttggtat gggcacagta
actacaacgg atacgatcca aaaaccggat 360gaggattact atggaagcaa
ggaagaccac ctttacaatg gtgatgacgc cttagttgat 420gaagttaagg
atatggaaat agcaagattg atgtctctag gtttatcaat tgaagaagcc
480actgagtttt acgaaaatga cgtaacttat gaaagatatt tggagatttt
aaagtcaaag 540caaaaggagc gcaacgatct agctataaga aagaaagaaa
gtggtataaa aatggaaaag 600tcaggattat ccaacattgt tggtacagat
agcaataatt tattcagcat ggccactgat 660tttttcaata agggtaagaa
actggtagac caatggacct ccttcccacc tgaggcaaat 720gatagactga
ataattactc aaaaactcat gataaggttg aggattatga tttgcctcaa
780gtaaacgact cacccaatag aattttgttt gaagataatg aagtcgtaga
gaacttacca 840cctgccgata atccggatca agatctttta actgatttcg
aaacaaagat tgatataaca 900aagaggacag cgcctgatgt ctcccactcc
tcctcaccga cttctggtat actaattgaa 960gaaaattcgc gaagaaatga
gcccctgata gaggatagtc ttctcgactt ttcagaagga 1020aatctcacca
atagtaaaag caatgaagat agcaccctct tcaatgaaaa cagcaacact
1080gactctacaa tacccatctc agatattgaa ttatcggggt ataacgaatt
taaggcgaaa 1140ggtactagtt tgttcaagaa cggggattat attaactcat
tacaagaata tgaaaagtct 1200ttaaatacat tgcctttaaa tcatccattg
aggatcattg cattatcaaa cattattgcc 1260tcgcaactga aaatcggtga
gtactctaag tccatagaaa actccagcat ggctttggaa 1320ttattcccat
caagcaaagc taagtggaag aataaaatct caaatagtga ccctgaaaga
1380tcatttaacg acatctggcc aaagattatg attaggcgtg ctgagtcttt
tgaacattta 1440gaaagtttca aaaaagcact agaaacatac caagagctga
ttaagaagaa tttttttgat 1500gataaaatca tgcagggaaa aagaagatgc
caagacttta ttaatcctcc ccctgttaaa 1560aaatccatgc ccgttaagaa
gaagacaacg acaacctcgc ctgcaacaaa aaaacagaac 1620ttaaccgctt
cttcttcaaa ttctccaatt tctgttgata gcacttcaga aataaaaaaa
1680cgggagctag aaaacgctaa actggcgcta tatgataaag tatttgagaa
aattagctcc 1740tggaaggatg gcaaagacga tgacattcgt catctgttag
caaatttatc cagcttacta 1800acatggtgca attggaagga tgtctctatg
caagatttgg ttatgcctaa gagggtcaaa 1860attacataca tgaaagctgt
agccaagaca catcctgata agataccaga gtccttgtcc 1920ctggaaaata
agatgattgc agagaatatt ttcagtactt taagtattgc ttgggataag
1980ttcaaactgc agaatgacat taactga 200777590PRTSaccharomyces
cerevisiae 77Met Lys Thr Cys Tyr Tyr Glu Leu Leu Gly Val Glu Thr
His Ala Ser1 5 10 15Asp Leu Glu Leu Lys Lys Ala Tyr Arg Lys Lys Ala
Leu Gln Tyr His 20 25 30Pro Asp Lys Asn Pro Asp Asn Val Glu Glu Ala
Thr Gln Lys Phe Ala 35 40 45Val Ile Arg Ala Ala Tyr Glu Val Leu Ser
Asp Pro Gln Glu Arg Ala 50 55 60Trp Tyr Asp Ser His Lys Glu Gln Ile
Leu Asn Asp Thr Pro Pro Ser65 70 75 80Thr Asp Asp Tyr Tyr Asp Tyr
Glu Val Asp Ala Thr Val Thr Gly Val 85 90 95Thr Thr Asp Glu Leu Leu
Leu Phe Phe Asn Ser Ala Leu Tyr Thr Lys 100 105 110Ile Asp Asn Ser
Ala Ala Gly Ile Tyr Gln Ile Ala Gly Lys Ile Phe 115 120 125Ala Lys
Leu Ala Lys Asp Glu Ile Leu Ser Gly Lys Arg Leu Gly Lys 130 135
140Phe Ser Glu Tyr Gln Asp Asp Val Phe Glu Gln Asp Ile Asn Ser
Ile145 150 155 160Gly Tyr Leu Lys Ala Cys Asp Asn Phe Ile Asn Lys
Thr Asp Lys Leu 165 170 175Leu Tyr Pro Leu Phe Gly Tyr Ser Pro Thr
Asp Tyr Glu Tyr Leu Lys 180 185 190His Phe Tyr Lys Thr Trp Ser Ala
Phe Asn Thr Leu Lys Ser Phe Ser 195 200 205Trp Lys Asp Glu Tyr Met
Tyr Ser Lys Asn Tyr Asp Arg Arg Thr Lys 210 215 220Arg Glu Val Asn
Arg Arg Asn Glu Lys Ala Arg Gln Gln Ala Arg Asn225 230 235 240Glu
Tyr Asn Lys Thr Val Lys Arg Phe Val Val Phe Ile Lys Lys Leu 245 250
255Asp Lys Arg Met Lys Glu Gly Ala Lys Ile Ala Glu Glu Gln Arg Lys
260 265 270Leu Lys Glu Gln Gln Arg Lys Asn Glu Leu Asn Asn Arg Arg
Lys Phe 275 280 285Gly Asn Asp Asn Asn Asp Glu Glu Lys Phe His Leu
Gln Ser Trp Gln 290 295 300Thr Val Lys Glu Glu Asn Trp Asp Glu Leu
Glu Lys Val Tyr Asp Asn305 310 315 320Phe Gly Glu Phe Glu Asn Ser
Lys Asn Asp Lys Glu Gly Glu Val Leu 325 330 335Ile Tyr Glu Cys Phe
Ile Cys Asn Lys Thr Phe Lys Ser Glu Lys Gln 340 345 350Leu Lys Asn
His Ile Asn Thr Lys Leu His Lys Lys Asn Met Glu Glu 355 360 365Ile
Arg Lys Glu Met Glu Glu Glu Asn Ile Thr Leu Gly Leu Asp Asn 370 375
380Leu Ser Asp Leu Glu Lys Phe Asp Ser Ala Asp Glu Ser Val Lys
Glu385 390 395 400Lys Glu Asp Ile Asp Leu Gln Ala Leu Gln Ala Glu
Leu Ala Glu Ile 405 410 415Glu Arg Lys Leu Ala Glu Ser Ser Ser Glu
Asp Glu Ser Glu Asp Asp 420 425 430Asn Leu Asn Ile Glu Met Asp Ile
Glu Val Glu Asp Val Ser Ser Asp 435 440 445Glu Asn Val His Val Asn
Thr Lys Asn Lys Lys Lys Arg Lys Lys Lys 450 455 460Lys Lys Ala Lys
Val Asp Thr Glu Thr Glu Glu Ser Glu Ser Phe Asp465 470 475 480Asp
Thr Lys Asp Lys Arg Ser Asn Glu Leu Asp Asp Leu Leu Ala Ser 485 490
495Leu Gly Asp Lys Gly Leu Gln Thr Asp Asp Asp Glu Asp Trp Ser Thr
500 505 510Lys Ala Lys Lys Lys Lys Gly Lys Gln Pro Lys Lys Asn Ser
Lys Ser 515 520 525Thr Lys Ser Thr Pro Ser Leu Ser Thr Leu Pro Ser
Ser Met Ser Pro 530 535 540Thr Ser Ala Ile Glu Val Cys Thr Thr Cys
Gly Glu Ser Phe Asp Ser545 550 555 560Arg Asn Lys Leu Phe Asn His
Val Lys Ile Ala Gly His Ala Ala Val 565 570 575Lys Asn Val Val Lys
Arg Lys Lys Val Lys Thr Lys Arg Ile 580 585
590781773DNASaccharomyces cerevisiae 78atgaagacct gctactatga
gcttttaggg gtcgaaacgc atgcttctga tcttgagtta 60aaaaaagctt accgtaaaaa
ggccctacaa tatcacccag ataaaaaccc agataatgtt 120gaagaagcca
cacaaaaatt tgctgtgatt cgagccgctt atgaagtact gtctgacccc
180caggaaagag catggtatga ctcacataag gaacaaattt taaatgatac
tccaccaagc 240actgatgatt actatgatta tgaggtagac gctacagtca
caggtgtcac aactgatgaa 300ttactcttat tttttaactc tgctctttat
actaaaatag acaactcagc tgctgggata 360tatcaaattg caggaaaaat
atttgccaag ttagctaaag atgagatttt aagtggtaag 420cgactgggga
aattttccga gtatcaagat gatgtattcg aacaggatat taatagtatt
480ggctatttga aagcctgcga taactttatt aacaagacgg ataaactttt
atatccttta 540tttggatatt cgccaacgga ttatgaatat ttgaaacatt
tctataagac ttggtcagcg 600ttcaatacct tgaaaagttt tagctggaaa
gacgagtaca tgtactctaa aaactatgac 660agaagaacca agagggaagt
taatagaaga aatgagaagg ctaggcaaca agctcgaaat 720gaatacaaca
aaaccgtgaa aaggtttgta gttttcataa aaaagctcga taaaagaatg
780aaagaaggtg caaaaattgc agaagaacag cgtaaactaa aagaacaaca
gaggaaaaat 840gagttaaata acagaagaaa gtttgggaac gacaacaatg
acgaagaaaa atttcattta 900caaagctggc aaacggtaaa agaagaaaac
tgggatgaac tggaaaaggt atatgataat 960tttggagaat tcgaaaattc
taagaatgat aaggaaggtg aagtattgat ttacgagtgt 1020tttatctgca
acaagacatt taagtcggaa aagcaattga aaaaccacat aaacactaaa
1080ctgcataaga aaaatatgga agagatacgg aaagaaatgg aagaggaaaa
cataacgctt 1140gggttggata atctctccga tctcgagaaa tttgattcag
cagatgaaag tgttaaagaa 1200aaagaagata ttgatctgca agcattgcaa
gctgaactcg ctgaaattga aagaaaactg 1260gcagaatcgt cttctgaaga
cgaaagtgaa gatgacaatc tcaacataga aatggatata 1320gaggtagaag
acgtcagttc ggatgaaaat gtacatgtga atacgaagaa taaaaagaaa
1380agaaaaaaga aaaaaaaagc aaaggttgac acagaaacag aggaatctga
atcgttcgat 1440gatactaaag acaaacggag taatgagttg gatgatcttt
tggcatcact aggagacaag 1500ggcttacaaa cggatgacga tgaagattgg
tctactaaag cgaaaaagaa aaagggcaaa 1560caacctaaaa agaattctaa
atccacaaaa agcactccgt ccttgtcgac tctaccgtcc 1620tctatgtctc
caacctccgc gatcgaggtg tgcactacat gcggagaatc atttgatagt
1680cgaaataagc tattcaacca cgtgaagata gcagggcatg cggcagtgaa
aaacgtagtg 1740aaaagaaaga aagtcaagac caaaagaata tag
177379583PRTSaccharomyces cerevisiae 79Met Ser Gln Val Ile Glu Pro
Gln Leu Asp Arg Thr Thr Tyr Tyr Ser1 5 10 15Ile Leu Gly Leu Thr Ser
Asn Ala Thr Ser Ser Glu Val His Lys Ser 20 25 30Tyr Leu Lys Leu Ala
Arg Leu Leu His Pro Asp Lys Thr Lys Ser Asp 35 40 45Lys Ser Glu Glu
Leu Phe Lys Ala Val Val His Ala His Ser Ile Leu 50 55 60Thr Asp Glu
Asp Gln Lys Leu Arg Tyr Asp Arg Asp Leu Lys Ile Lys65 70 75 80Gly
Leu His Thr Tyr Gln Pro Lys Lys Asn Cys His Ile Phe Lys Thr 85 90
95Lys Ala Lys Glu Ser Gln Gly Ala Ser Pro Thr Leu Gly Gln Ser Glu
100 105 110Ala Tyr His Arg Gln Asn Lys Pro Tyr Glu Gln Gln Pro Tyr
Gly Phe 115 120 125Gly Val Gly Lys Lys Met Thr Ser Ser Ser Lys Ser
Lys Val Pro Ile 130 135 140Phe Lys Ser Phe Asn Leu Lys Ser Tyr Gln
Arg Asn His Tyr Tyr Ser145 150 155 160Ser Lys Lys Glu Arg Lys His
Gly Ser Pro Asp Ile Asp Ser Leu Phe 165 170 175His Glu Thr Asn Gly
Ala Ser Lys Val Arg Met Thr Asp Ala Gly Lys 180 185 190Met Asp Thr
Asn Ser Gln Phe Gln Glu Ile Trp Glu Ile Leu Gly Lys 195 200 205Asn
Ala Tyr Thr His Lys Ser Tyr Ser Glu Asp Pro Asn Ser Cys Leu 210 215
220Gly Ser Ala Leu Ser Asp His Glu Glu Glu Glu Glu Ala Gly Lys
Gln225 230 235 240Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln
His Tyr Gly Met 245 250 255Thr Ser Lys Ser Ser Ser Pro Asp Glu Glu
Lys Lys Asn Asn Lys Glu 260 265 270Pro Lys Arg Glu Ser Arg Val Ser
Pro Glu Glu Asn Gly Glu Glu Glu 275 280 285Thr Gly His Lys Gln Phe
Lys Leu Pro Lys Thr Ser Thr Phe Ser Ser 290 295 300Gly Ser His Asp
Ser Asn Leu Gln Ser Pro Phe Tyr Asn His Glu Tyr305 310 315 320Arg
His Tyr Ala Arg Ser Lys Phe Glu Cys Lys Asn Gln Phe Arg Lys 325 330
335Ser Val Ser Pro Ile Lys Glu Ile Pro Ala Thr Thr Ser Ala Asn Glu
340 345 350Gly Trp Asn Ile Leu Arg Asp Ile Ile Glu Lys Leu Asn Ile
Ser Asn 355 360 365Val Asp Asp Arg Asn Lys Asp Leu Leu Phe Arg Arg
Asp Glu Ile Gly 370 375 380Asp Lys Asn His Ser Asp Ser Ile Asp Ile
Glu Asn Leu Ser Ile Lys385 390 395 400Glu Pro Lys Gly Met Lys Arg
Arg Lys Lys Asp Asp Ile Ser Leu Glu 405 410 415Glu Leu Phe Gln Ser
Leu Pro Arg Glu Lys Asp Tyr Phe Met Met Asp 420 425 430Ala Ile Asn
Asp Ser Leu Glu Ser Ile Asn Leu Phe Lys Lys Pro Lys 435 440 445Thr
Thr Gln Ser His Glu Gln Gly Gly Thr Phe Ala Gln Ala Glu Ser 450 455
460Asn Arg Ala Lys Phe Lys Pro Leu Leu Glu Gln Cys Gly Ile Thr
Pro465 470 475 480Glu Ile Leu Asp Leu Glu Ile Pro Glu Ile Pro Glu
Phe Asp Ala Val 485 490 495Ala Asp Leu Glu Thr Leu Lys Leu Asn Val
Gln Leu Phe Asn Asn Gln 500 505 510Cys Asn Lys Leu Lys Glu Thr Ile
His Gln Val Ser Leu Gln Arg Leu 515 520 525Arg Ala Asp Thr Gln Phe
Ser Asp Met Leu Thr Gln Lys Gln Ser Ile 530 535 540Met Val Trp Lys
Thr Tyr Leu Glu Phe Asp Lys Ser Leu Met Asp Lys545 550 555 560Leu
Asn Ile Leu Gln Glu Arg Gln Met Gln Val Ile Lys Ile Phe Ser 565 570
575Glu Arg Cys Asp Gly Lys Val 580801752DNASaccharomyces cerevisiae
80atgtcacagg taatagaacc acaattagat agaacaacct attattccat attaggcttg
60acatcaaatg cgacttcctc cgaagtacat aaatcatatc taaaactggc cagattactt
120cacccagata aaacaaaatc tgataagtct gaggaattat tcaaagctgt
ggtgcatgca 180cattcaattt taactgatga agatcaaaaa cttcgatatg
atcgagattt gaaaatcaaa 240ggtttacaca cttaccagcc gaagaaaaac
tgtcatattt tcaagaccaa ggcaaaggaa 300tcacaagggg ctagtcccac
acttggtcaa tcagaagctt atcataggca aaataaacct 360tatgagcaac
agccctacgg tttcggtgta ggcaaaaaaa tgacctcaag ctctaagagt
420aaggttccga tattcaagtc cttcaattta aaaagctacc aacgaaacca
ctattattca 480tccaaaaagg aaaggaaaca tggaagtcct gatattgatt
ctttgttcca tgaaaccaat 540ggagcctcaa aagtaagaat gactgatgcc
ggtaaaatgg atacgaactc tcagttccaa 600gaaatatggg aaatattggg
taaaaatgcg tacacacata aatcttactc tgaagatcca 660aattcatgtt
tgggatcagc actaagcgat catgaagaag aagaagaagc aggaaaacaa
720caacagcaac agcagcaaca acagcaacag cagcaacatt atggaatgac
gtcgaagtct 780agcagtcctg atgaagaaaa aaaaaataat aaagaaccga
aaagggaaag cagagtctct 840ccagaggaaa atggcgaaga agaaacggga
cacaaacaat ttaaattgcc caagaccagt 900actttttcta gtggatccca
tgattcaaat ttgcaatctc ctttttacaa tcatgagtat 960cgacattacg
caagaagtaa attcgaatgc aagaatcagt ttagaaagtc agtttctccc
1020attaaagaga tacctgcaac aactagtgcc aatgaaggat ggaacatttt
gagagacatt 1080attgaaaaac tcaatataag caatgtagac gatcgaaata
aagacttgct gtttcgtcgg 1140gatgaaatag gtgataaaaa tcacagcgac
tcaatcgaca tagaaaattt atctatcaaa 1200gaacctaaag ggatgaaaag
gagaaagaaa gatgatatat ctttagaaga attgttccaa 1260tctttaccaa
gagaaaaaga ttattttatg atggatgcaa ttaatgactc gttagaatca
1320atcaatcttt ttaaaaagcc gaagaccact cagagtcacg aacaaggtgg
aacttttgcc 1380caagcagaaa gtaatcgtgc aaaattcaaa ccgttactag
aacagtgtgg aattacaccc 1440gagatcttag atttggaaat accagagatt
ccggaatttg atgcagtggc tgaccttgaa 1500acattgaagc ttaacgtgca
gctgtttaat aaccaatgta acaaacttaa agaaacaata 1560catcaagtat
cattacagcg cctgagagca gatacgcagt tcagtgatat gttaacccaa
1620aagcaaagta ttatggtttg gaaaacatac ctagaatttg ataaaagttt
aatggacaaa 1680ttgaacatct tacaagaaag acagatgcag gtcattaaaa
ttttttccga aagatgtgac 1740ggtaaagtat aa 175281172PRTSaccharomyces
cerevisiae 81Met Ser Leu Val Asn Ser Leu Thr His Tyr Glu Ile Leu
Arg Ile Pro1 5 10 15Ser Asp Ala Thr Gln Asp Glu Ile Lys Lys Ala Tyr
Arg Asn Arg Leu 20 25 30Leu Asn Thr His Pro Asp Lys Leu Ser Lys Ser
Ile His Asp Thr Val 35 40 45Ser Asn Val Thr Ile Asn Lys Ile Gln Asp
Ala Tyr Lys Ile Leu Ser 50 55 60Asn Ile Lys Thr Arg Arg Glu Tyr Asp
Arg Leu Ile Leu Glu Asn Tyr65 70 75 80Lys Arg Gln Gly Phe His Asn
Cys Gly Asp Gly Leu Asp Glu Phe Ser 85 90 95Leu Asp Asp Phe Ser Phe
Asp Glu Asp Lys Leu Glu Phe Met Met Asn 100 105 110Cys Pro Arg Cys
Gln Phe Val Gly Gly Phe His Phe Ser Glu Ser Leu 115 120 125Leu Asp
Glu Cys Ile Asp Asn Val Asp Ala Met Glu Arg Ser His Ser 130 135
140Gly Tyr Gln Leu Leu Thr Gln Cys Ser Ala Cys Ser Leu Trp Leu
Lys145 150 155 160Val Asn Phe Asp Ile Glu Glu Glu Gln Glu Gly Gln
165 17082519DNASaccharomyces cerevisiae 82atgtcattgg tgaattcgtt
aacacactac gaaattttaa gaattccatc ggatgcaaca 60caagatgaaa tcaaaaaggc
atataggaat cggttactaa atacgcaccc cgataaactt 120tctaaaagca
tacatgatac ggttagcaac gtcacaatca ataagattca agatgcttat
180aaaatactat cgaatataaa aactcgtcgc gaatatgata ggttgatcct
tgaaaactat 240aaacgccaag gatttcataa ttgtggtgat gggctggatg
aattttcctt agacgatttc 300tcatttgatg aagataagct ggagtttatg
atgaattgtc ctcgctgtca atttgttggt 360ggttttcatt ttagtgagag
tttgttagat gaatgcattg ataatgtaga cgctatggaa 420cggagtcatt
ctggttatca attattaacc caatgtagcg catgcagctt atggctgaag
480gttaattttg acatcgagga agagcaagaa ggacaataa
51983391PRTSaccharomyces cerevisiae 83Met Val Lys Glu Thr Glu Tyr
Tyr Asp Ile Leu Gly Ile Lys Pro Glu1 5 10 15Ala Thr Pro Thr Glu Ile
Lys Lys Ala Tyr Arg Arg Lys Ala Met Glu 20 25 30Thr His Pro Asp Lys
His Pro Asp Asp Pro Asp Ala Gln Ala Lys Phe 35
40 45Gln Ala Val Gly Glu Ala Tyr Gln Val Leu Ser Asp Pro Gly Leu
Arg 50 55 60Ser Lys Tyr Asp Gln Phe Gly Lys Glu Asp Ala Val Pro Gln
Gln Gly65 70 75 80Phe Glu Asp Ala Ser Glu Tyr Phe Thr Ala Ile Phe
Gly Gly Asp Gly 85 90 95Phe Lys Asp Trp Ile Gly Glu Phe Ser Leu Phe
Lys Glu Leu Asn Glu 100 105 110Ala Thr Glu Met Phe Gly Lys Glu Asp
Glu Glu Gly Thr Ala Ala Thr 115 120 125Glu Thr Glu Lys Ala Asp Glu
Ser Thr Asp Gly Gly Met Val Lys His 130 135 140Asp Thr Asn Lys Ala
Glu Ser Leu Lys Lys Asp Lys Leu Ser Lys Glu145 150 155 160Gln Arg
Glu Lys Leu Met Glu Met Glu Lys Lys Arg Arg Glu Asp Met 165 170
175Met Lys Gln Val Asp Glu Leu Ala Glu Lys Leu Asn Glu Lys Ile Ser
180 185 190Arg Tyr Leu Ile Ala Val Lys Ser Asn Asn Leu Glu Glu Phe
Thr Arg 195 200 205Lys Leu Asp Gln Glu Ile Glu Asp Leu Lys Leu Glu
Ser Phe Gly Leu 210 215 220Glu Leu Leu Tyr Leu Leu Ala Arg Val Tyr
Lys Thr Lys Ala Asn Asn225 230 235 240Phe Ile Met Ser Lys Lys Thr
Tyr Gly Ile Ser Lys Ile Phe Thr Gly 245 250 255Thr Arg Asp Asn Ala
Arg Ser Val Lys Ser Ala Tyr Asn Leu Leu Ser 260 265 270Thr Gly Leu
Glu Ala Gln Lys Ala Met Glu Lys Met Ser Glu Val Asn 275 280 285Thr
Asp Glu Leu Asp Gln Tyr Glu Arg Ala Lys Phe Glu Ser Thr Met 290 295
300Ala Gly Lys Ala Leu Gly Val Met Trp Ala Met Ser Lys Phe Glu
Leu305 310 315 320Glu Arg Lys Leu Lys Asp Val Cys Asn Lys Ile Leu
Asn Asp Lys Lys 325 330 335Val Pro Ser Lys Glu Arg Ile Ala Lys Ala
Lys Ala Met Leu Phe Ile 340 345 350Ala His Lys Phe Ala Ser Ala Arg
Arg Ser Pro Glu Glu Ala Glu Glu 355 360 365Ala Arg Val Phe Glu Glu
Leu Ile Leu Gly Glu Gln Glu Lys Glu His 370 375 380Lys Lys His Thr
Val Ala Arg385 390841176DNASaccharomyces cerevisiae 84atggtaaagg
agacggagta ttatgatatt ttgggcatca agcctgaggc cacgcccact 60gaaatcaaaa
aggcctatcg tagaaaggct atggaaacac atccggacaa gcatcctgat
120gacccagatg ctcaagcaaa gtttcaagcc gtaggcgagg cctaccaagt
cttaagtgat 180ccagggcttc gttccaagta tgaccagttt ggtaaggagg
atgctgttcc tcagcaagga 240tttgaagatg cttctgaata ctttacagca
atattcggtg gtgatggctt caaagattgg 300attggagaat tttctttgtt
caaagagcta aacgaggcaa cagaaatgtt tggaaaggaa 360gatgaggagg
gtacagcagc cactgaaacc gaaaaagcag atgagagcac tgatggtgga
420atggttaagc atgacactaa taaagctgaa tctttgaaaa aagataaatt
atcgaaggag 480caaagagaga agctaatgga aatggagaaa aaaagacggg
aagatatgat gaaacaagtc 540gacgagttgg cagaaaaact gaacgaaaaa
atctctaggt acttaattgc tgtgaagtcc 600aataacttgg aggaatttac
gcgaaaacta gatcaagaaa tcgaggattt gaaattagaa 660agttttggtc
tagagttatt gtatttattg gccagggttt acaagacaaa agcgaataat
720tttatcatgt ccaagaagac ttacggaatt tctaaaatat tcactggtac
acgcgacaat 780gctagatctg ttaaatcagc atacaattta ttgtctacag
gcttagaagc tcaaaaagcc 840atggaaaaaa tgagtgaagt caatactgac
gaactagacc aatatgaacg tgccaaattt 900gagtccacaa tggctggtaa
ggcacttggt gtcatgtggg ctatgtcgaa atttgaactg 960gaaagaaaac
taaaagacgt ttgcaataag attctaaacg ataaaaaggt cccttccaag
1020gaacgtattg caaaggcaaa agcaatgctg tttattgccc acaagtttgc
cagtgctaga 1080aggtcaccag aagaagctga agaagctaga gtttttgaag
agctaatcct aggtgagcag 1140gagaaggaac acaaaaaaca tactgtggcc agataa
117685283PRTSaccharomyces cerevisiae 85Met Pro Gly His Glu Leu Glu
Asp Val Ile Asn Gln Arg Leu Asn Leu1 5 10 15Tyr Asp Val Leu Glu Leu
Pro Thr Pro Leu Asp Val His Thr Ile Tyr 20 25 30Asp Asp Leu Pro Gln
Ile Lys Arg Lys Tyr Arg Thr Leu Ala Leu Lys 35 40 45Tyr His Pro Asp
Lys His Pro Asp Asn Pro Ser Ile Ile His Lys Phe 50 55 60His Leu Leu
Ser Thr Ala Thr Asn Ile Leu Thr Asn Ala Asp Val Arg65 70 75 80Pro
His Tyr Asp Arg Trp Leu Ile Glu Phe Leu Arg Lys Thr Asn Asp 85 90
95Ile Glu Arg Asn Lys Leu Ile Gln Lys Leu Glu Glu Ser Glu Ser Ser
100 105 110Thr Ile Pro Thr Thr Thr Pro His Pro Asp Leu Leu Gln Ile
Gln Arg 115 120 125His Gly Glu Leu Leu Arg Lys Leu Lys His Phe Asn
Leu Pro Tyr Gly 130 135 140Asp Trp Lys His Leu Asn Thr Gln Asp Gln
Glu Asn Ala Ser Gln His145 150 155 160Pro Tyr Tyr Asp Cys Ser Thr
Leu Arg Ile Val Leu Asp Asn Phe Leu 165 170 175Gln Ser Asn Asn Lys
Ser Asn Cys Leu Ser His Leu Arg Asn Gln Val 180 185 190Phe Ile Thr
Leu Ser Ala Asn Glu Ile Tyr Asp Ile Tyr Phe Ser Glu 195 200 205Arg
Asn Asn Tyr Ser Lys Asp Asp Ser Ile Ile Ile Tyr Thr Val Phe 210 215
220Asp Thr Pro Ile Thr Ala Gln His Val Phe Arg Asn Trp Ser Ser
Gly225 230 235 240Asn Leu Ile Pro Thr Val Lys Asp Ile Ser Pro Leu
Ile Pro Leu His 245 250 255Tyr Tyr Ser Asp Phe Asn Leu Glu Thr Glu
Leu Asn Asp Asp Ile Ala 260 265 270Arg Leu Val Ser Asn Glu Pro Ile
Leu Leu Asp 275 28086852DNASaccharomyces cerevisiae 86atgccaggac
acgaattgga agacgtaata aatcaacgtt tgaacctata tgatgtatta 60gaattaccga
cccccctgga cgtccatacc atctacgatg atttgcccca aattaaacgc
120aaatacagga cccttgccct gaagtatcat cctgacaaac acccggacaa
tccatcaatt 180atacacaaat tccacttatt atcgaccgca actaatatcc
tcaccaatgc agacgtgaga 240ccccattacg accgctggtt aattgagttc
ctacggaaaa caaacgacat tgaaagaaat 300aaacttatac aaaagctgga
agaatctgaa tcgagtacga tacccaccac cacaccacat 360cctgatttat
tgcaaatcca acgccacggc gagctactca ggaaactaaa acatttcaac
420ttgccctatg gtgactggaa acatctcaac acacaagacc aagaaaatgc
ttcgcaacat 480ccgtattacg attgctctac tttgagaatt gtccttgaca
acttcctgca atcaaataat 540aaatcaaact gcttatctca tttgcgcaat
caagtattca tcacgctaag tgctaatgaa 600atctacgaca tctacttctc
tgaaagaaac aactactcga aggatgattc aatcatcata 660tatactgtat
tcgatactcc catcacagcg cagcacgtat tccgaaactg gtcaagtggg
720aacctcatac ccacggtcaa ggatatttcg cccttgatcc cgctacatta
ctactctgat 780tttaatttgg agacggaact gaatgacgat attgcaagac
tggtctctaa tgaacctatc 840ctactcgact ag 85287168PRTSaccharomyces
cerevisiae 87Met Ser Ser Gln Ser Asn Thr Gly Asn Ser Ile Glu Ala
Pro Gln Leu1 5 10 15Pro Ile Pro Gly Gln Thr Asn Gly Ser Ala Asn Val
Thr Val Asp Gly 20 25 30Ala Gly Val Asn Val Gly Ile Gln Asn Gly Ser
Gln Gly Gln Lys Thr 35 40 45Gly Met Asp Leu Tyr Phe Asp Gln Ala Leu
Asn Tyr Met Gly Glu His 50 55 60Pro Val Ile Thr Gly Phe Gly Ala Phe
Leu Thr Leu Tyr Phe Thr Ala65 70 75 80Gly Ala Tyr Lys Ser Ile Ser
Lys Gly Leu Asn Gly Gly Lys Ser Thr 85 90 95Thr Ala Phe Leu Lys Gly
Gly Phe Asp Pro Lys Met Asn Ser Lys Glu 100 105 110Ala Leu Gln Ile
Leu Asn Leu Thr Glu Asn Thr Leu Thr Lys Lys Lys 115 120 125Leu Lys
Glu Val His Arg Lys Ile Met Leu Ala Asn His Pro Asp Lys 130 135
140Gly Gly Ser Pro Phe Leu Ala Thr Lys Ile Asn Glu Ala Lys Asp
Phe145 150 155 160Leu Glu Lys Arg Gly Ile Ser Lys
16588507DNASaccharomyces cerevisiae 88atgagttctc aaagtaatac
tggtaattct attgaggcac cacaactacc cattcctggt 60caaactaatg gctctgcgaa
cgttactgtt gatggagctg gtgttaatgt cggtatccag 120aatggttcgc
agggtcaaaa gaccggaatg gacctttatt ttgatcaagc tttgaactac
180atgggagaac atcctgtgat aacaggtttt ggggcctttt taactttata
ttttacagcc 240ggtgcatata aatcaatatc gaagggactt aacggtggaa
aatccactac tgccttcttg 300aaaggcggat ttgacccgaa aatgaattct
aaagaggctc tacagatttt gaatttgaca 360gaaaatacat tgactaaaaa
aaagttgaaa gaggttcata ggaaaattat gttagctaat 420catcctgaca
aaggtggttc tccatttttg gccactaaga taaacgaagc taaggacttt
480ttggaaaaaa ggggtattag caaataa 50789184PRTSaccharomyces
cerevisiae 89Met Leu Lys Tyr Leu Val Gln Arg Arg Phe Thr Ser Thr
Phe Tyr Glu1 5 10 15Leu Phe Pro Lys Thr Phe Pro Lys Lys Leu Pro Ile
Trp Thr Ile Asp 20 25 30Gln Ser Arg Leu Arg Lys Glu Tyr Arg Gln Leu
Gln Ala Gln His His 35 40 45Pro Asp Met Ala Gln Gln Gly Ser Glu Gln
Ser Ser Thr Leu Asn Gln 50 55 60Ala Tyr His Thr Leu Lys Asp Pro Leu
Arg Arg Ser Gln Tyr Met Leu65 70 75 80Lys Leu Leu Arg Asn Ile Asp
Leu Thr Gln Glu Gln Thr Ser Asn Glu 85 90 95Val Thr Thr Ser Asp Pro
Gln Leu Leu Leu Lys Val Leu Asp Ile His 100 105 110Asp Glu Leu Ser
Gln Met Asp Asp Glu Ala Gly Val Lys Leu Leu Glu 115 120 125Lys Gln
Asn Lys Glu Arg Ile Gln Asp Ile Glu Ala Gln Leu Gly Gln 130 135
140Cys Tyr Asn Asp Lys Asp Tyr Ala Ala Ala Val Lys Leu Thr Val
Glu145 150 155 160Leu Lys Tyr Trp Tyr Asn Leu Ala Lys Ala Phe Lys
Asp Trp Ala Pro 165 170 175Gly Lys Gln Leu Glu Met Asn His
18090555DNASaccharomyces cerevisiae 90atgttgaaat acttggttca
acgaagattc acttctacat tttacgagct gttcccaaag 60accttcccca aaaagctacc
catttggact atcgatcaat ccagattaag gaaggagtat 120aggcaattac
aagcacagca ccatccagac atggcccaac aaggtagtga acagtcatca
180actcttaatc aagcttacca tactctcaaa gatcccctta gaaggtcaca
atatatgcta 240aaactcttgc gcaatatcga tttgacgcaa gaacagacct
caaatgaagt aactaccagt 300gatccacagt tactattgaa agttctagac
atccatgatg aattatccca gatggacgac 360gaagctggtg tgaagctgct
tgaaaagcaa aacaaggaaa gaattcaaga tattgaagcc 420cagttgggac
aatgctacaa tgacaaggat tacgccgccg cagtgaagtt gaccgtggag
480ctaaagtact ggtacaactt ggccaaggca ttcaaagact gggctccagg
aaaacaattg 540gaaatgaatc actaa 55591301PRTSaccharomyces cerevisiae
91Met Leu His His Lys Phe Val Tyr Pro Phe Leu Phe Lys Trp His Leu1
5 10 15Ser Cys Val Glu Lys Cys Pro Pro Gln Ile Thr Phe Ile Ala Lys
Tyr 20 25 30Ala Thr Ala Asn Asp Lys Asn Gly Asn Arg Lys Leu Thr Ile
Arg Asp 35 40 45Glu Gln Trp Pro Glu Leu Ala Asp Pro Thr Pro Tyr Asp
Ile Phe Gly 50 55 60Ile Pro Lys Ala Gly Ser Gly Asn Pro Lys Leu Asp
Lys Lys Ser Leu65 70 75 80Lys Lys Lys Tyr His Arg Tyr Val Lys Leu
Tyr His Pro Asp His Ser 85 90 95Asp Asn Ile Gln Ile Phe Ser Ser Glu
Lys Val Thr Asn Ser Asp Ser 100 105 110Lys Ser Pro Leu Leu Leu Thr
Ser Ser Glu Lys Leu His Arg Phe Lys 115 120 125Val Ile Ser Gln Ala
Tyr Asp Ile Leu Cys Asp Pro Lys Lys Lys Ile 130 135 140Val Tyr Asp
Thr Thr Arg Gln Gly Trp Thr Thr Ser Tyr Ser Pro Arg145 150 155
160Ser Asn Val Asn Thr Glu Asn Tyr Gln Tyr Ala Gly Ser Tyr Gly Tyr
165 170 175His Ser Asn Ala Gln Tyr Glu Tyr Trp Asn Ala Gly Thr Trp
Glu Asp 180 185 190Ala Asn Ser Met Lys Asn Glu Arg Ile Gln Glu Asn
Ile Asn Pro Trp 195 200 205Thr Val Ile Gly Ile Ile Cys Gly Leu Ala
Ile Cys Ile Glu Gly Thr 210 215 220Ala Leu Leu Ala Lys Ile Gln Glu
Ser Leu Ser Lys Ala Glu Phe Thr225 230 235 240His Asp Glu Ser Gly
Leu His Leu Ile Gln Ser Tyr Thr Asn Tyr Gly 245 250 255Leu Asp Thr
Asp Lys Phe Ser Arg Leu Arg Arg Phe Leu Trp Phe Arg 260 265 270Thr
Trp Gly Leu Tyr Lys Ser Lys Glu Asp Leu Asp Arg Glu Ala Lys 275 280
285Ile Asn Glu Glu Met Ile Arg Lys Leu Lys Ala Ala Lys 290 295
30092906DNASaccharomyces cerevisiae 92atgctacacc ataagttcgt
atacccattt ttattcaagt ggcacttatc atgtgtagaa 60aagtgtcccc cacaaatcac
ttttatagct aagtatgcta cagcgaacga taaaaatggc 120aatagaaaac
ttacgataag ggatgaacaa tggcctgagt tggcagatcc aactccctat
180gatatttttg gcattccaaa ggccggatct ggaaatccta aactggacaa
gaagtcgtta 240aaaaaaaaat atcatcgtta tgtaaaattg taccaccctg
accattccga taacattcaa 300atatttagct cagaaaaggt taccaacagt
gatagtaaat caccgctgct gctaacatca 360agcgaaaaac tacatagatt
taaagtcatc tctcaagcat atgatattct ttgtgaccca 420aagaaaaaga
tcgtatatga cacaacgagg caaggctgga ccacatcgta ttcaccacgt
480tctaacgtta atactgaaaa ttaccaatat gccggctctt atggctacca
ctctaacgcg 540cagtatgaat actggaacgc tgggacttgg gaagacgcaa
atagcatgaa aaacgaaaga 600attcaagaaa acatcaaccc atggaccgtt
attggcataa tttgtggcct agctatatgc 660atcgaaggga ctgcgttgtt
agccaaaatc caggagtctc tgagcaaggc cgaatttact 720catgacgaaa
gtggattaca tttgattcag tcatacacga attatggtct tgatactgac
780aaattttcca gattgaggcg gttcttatgg tttagaactt ggggacttta
caagtcgaaa 840gaggatttag atagagaagc caagatcaat gaagaaatga
tacgcaaact gaaagcagct 900aaatga 90693224PRTSaccharomyces cerevisiae
93Met Ser Phe Thr Glu Asp Gln Glu Lys Ile Ala Leu Glu Ile Leu Ser1
5 10 15Lys Asp Lys His Glu Phe Tyr Glu Ile Leu Lys Val Asp Arg Lys
Ala 20 25 30Thr Asp Ser Glu Ile Lys Lys Ala Tyr Arg Lys Leu Ala Ile
Lys Leu 35 40 45His Pro Asp Lys Asn Ser His Pro Lys Ala Gly Glu Ala
Phe Lys Val 50 55 60Ile Asn Arg Ala Phe Glu Val Leu Ser Asn Glu Glu
Lys Arg Ser Ile65 70 75 80Tyr Asp Arg Ile Gly Arg Asp Pro Asp Asp
Arg Gln Met Pro Ser Arg 85 90 95Gly Ala Ala Ser Gly Phe Arg Gly Ser
Ala Gly Gly Ser Pro Met Gly 100 105 110Gly Gly Phe Glu Asp Met Phe
Phe Asn Ser Arg Phe Gly Gly Gln Arg 115 120 125Ala Gly Pro Pro Glu
Asp Ile Phe Asp Phe Leu Phe Asn Ala Gly Gly 130 135 140Ser Pro Phe
Gly Ala Ser Pro Phe Gly Pro Ser Ala Ser Thr Phe Ser145 150 155
160Phe Gly Gly Pro Gly Gly Phe Arg Val Tyr Thr Asn Asn Arg Gly Gly
165 170 175Ser Pro Phe Met Arg Gln Gln Pro Arg Ser Arg Gln Gln Gln
Gln Gln 180 185 190Ala Glu Glu Asn Ala Val Asn Ser Gln Leu Lys Asn
Met Leu Val Leu 195 200 205Phe Ile Ile Phe Ile Val Leu Pro Met Ile
Lys Asp Tyr Leu Phe Ser 210 215 22094675DNASaccharomyces cerevisiae
94atgtctttca ctgaggatca agaaaaaatc gcgctagaaa tactgtcaaa agacaagcat
60gagttttacg aaattttgaa ggtagatagg aaagccacag atagtgagat caagaaggca
120tacagaaaac tagcaatcaa attgcatcct gataaaaact ctcatccaaa
agcgggagaa 180gctttcaaag taattaatag ggcatttgaa gtactaagca
atgaggaaaa gcgcagtatt 240tatgacagga taggtaggga tcctgacgat
agacaaatgc catccagagg tgctgcttca 300gggttccgag gaagtgcagg
tgggtctcca atgggtggcg gatttgaaga catgtttttc 360aattcacgtt
tcggtggtca aagagctgga ccaccagagg acatattcga ctttttgttc
420aacgcaggcg gcagcccatt cggcgcttca ccatttgggc cttctgcttc
cactttttca 480tttggaggcc ccggtggttt cagagtttat actaataatc
gtggtggctc accgttcatg 540cgtcaacaac cccgctcaag acagcagcaa
caacaagcag aagaaaatgc agtgaattcg 600caattaaaaa atatgctcgt
tcttttcatc atctttattg ttcttcctat gattaaagat 660tacctgttta gttaa
67595295PRTSaccharomyces cerevisiae 95Met Asn Gly Tyr Trp Lys Pro
Ala Leu Val Val Leu Gly Leu Val Ser1 5 10 15Leu Ser Tyr Ala Phe Thr
Thr Ile Glu Thr Glu Ile Phe Gln Leu Gln 20 25 30Asn Glu Ile Ser Thr
Lys Tyr Gly Pro Asp Met Asn Phe Tyr Lys Phe 35 40 45Leu Lys Leu Pro
Lys Leu Gln Asn Ser Ser Thr Lys Glu Ile Thr Lys 50 55 60Asn Leu Arg
Lys Leu Ser Lys Lys Tyr His Pro Asp Lys Asn Pro Lys65 70 75 80Tyr
Arg Lys Leu Tyr Glu Arg Leu Asn Leu Ala Thr Gln Ile Leu Ser 85 90
95Asn Ser Ser Asn Arg Lys Ile Tyr Asp Tyr Tyr Leu Gln Asn Gly Phe
100
105 110Pro Asn Tyr Asp Phe His Lys Gly Gly Phe Tyr Phe Ser Arg Met
Lys 115 120 125Pro Lys Thr Trp Phe Leu Leu Ala Phe Ile Trp Ile Val
Val Asn Ile 130 135 140Gly Gln Tyr Ile Ile Ser Ile Ile Gln Tyr Arg
Ser Gln Arg Ser Arg145 150 155 160Ile Glu Asn Phe Ile Ser Gln Cys
Lys Gln Gln Asp Asp Thr Asn Gly 165 170 175Leu Gly Val Lys Gln Leu
Thr Phe Lys Gln His Glu Lys Asp Glu Gly 180 185 190Lys Ser Leu Val
Val Arg Phe Ser Asp Val Tyr Val Val Glu Pro Asp 195 200 205Gly Ser
Glu Thr Leu Ile Ser Pro Asp Thr Leu Asp Lys Pro Ser Val 210 215
220Lys Asn Cys Leu Phe Trp Arg Ile Pro Ala Ser Val Trp Asn Met
Thr225 230 235 240Phe Gly Lys Ser Val Gly Ser Ala Gly Lys Glu Glu
Ile Ile Thr Asp 245 250 255Ser Lys Lys Tyr Asp Gly Asn Gln Thr Lys
Lys Gly Asn Lys Val Lys 260 265 270Lys Gly Ser Ala Lys Lys Gly Gln
Lys Lys Met Glu Leu Pro Asn Gly 275 280 285Lys Val Ile Tyr Ser Arg
Lys 290 29596888DNASaccharomyces cerevisiae 96atgaacggtt actggaaacc
tgcgttggtt gtcctgggat tggtatctct atcatatgct 60tttaccacca ttgaaacaga
aattttccaa ttacaaaatg aaataagtac gaaatatggc 120ccagatatga
acttctacaa gttcttgaag ttacctaaac tgcagaattc tagtacaaag
180gagattacaa aaaacttaag aaagctatcc aagaagtacc atccggataa
gaaccctaaa 240taccgtaaat tgtatgaaag gttaaacctc gctactcaaa
ttctttcaaa cagctctaat 300cgtaagattt atgattatta tctacagaat
ggctttccaa actatgattt ccataagggt 360ggtttttatt tttccagaat
gaagcctaag acttggttcc tgctggcctt tatttggata 420gtcgttaata
ttgggcagta tatcatttct attattcaat atcgttctca aagatcaaga
480attgaaaact tcatcagtca gtgtaaacaa caggatgata ccaatggact
aggcgtaaaa 540caactaacgt ttaaacaaca tgaaaaggat gagggtaaaa
gtttggttgt aaggtttagc 600gatgtctatg ttgtagagcc tgatggaagt
gaaacactaa tttcgccaga taccttggat 660aaaccttcag taaagaactg
tttgttttgg agaatacctg cttcggtttg gaacatgacg 720tttggcaaat
ctgttggtag cgcaggaaaa gaagaaataa taacggatag taaaaagtat
780gatggtaacc aaacaaaaaa ggggaacaaa gtaaaaaagg gttctgcaaa
gaaaggccaa 840aagaaaatgg aattgcctaa cggtaaagtg atctattcac gtaaatga
88897228PRTSaccharomyces cerevisiae 97Met Arg Ala Phe Ser Ala Ala
Thr Val Arg Ala Thr Thr Arg Lys Ser1 5 10 15Phe Ile Pro Met Ala Pro
Arg Thr Pro Phe Val Thr Pro Ser Phe Thr 20 25 30Lys Asn Val Gly Ser
Met Arg Arg Met Arg Phe Tyr Ser Asp Glu Ala 35 40 45Lys Ser Glu Glu
Ser Lys Glu Asn Asn Glu Asp Leu Thr Glu Glu Gln 50 55 60Ser Glu Ile
Lys Lys Leu Glu Ser Gln Leu Ser Ala Lys Thr Lys Glu65 70 75 80Ala
Ser Glu Leu Lys Asp Arg Leu Leu Arg Ser Val Ala Asp Phe Arg 85 90
95Asn Leu Gln Gln Val Thr Lys Lys Asp Ile Gln Lys Ala Lys Asp Phe
100 105 110Ala Leu Gln Lys Phe Ala Lys Asp Leu Leu Glu Ser Val Asp
Asn Phe 115 120 125Gly His Ala Leu Asn Ala Phe Lys Glu Glu Asp Leu
Gln Lys Ser Lys 130 135 140Glu Ile Ser Asp Leu Tyr Thr Gly Val Arg
Met Thr Arg Asp Val Phe145 150 155 160Glu Asn Thr Leu Arg Lys His
Gly Ile Glu Lys Leu Asp Pro Leu Gly 165 170 175Glu Pro Phe Asp Pro
Asn Lys His Glu Ala Thr Phe Glu Leu Pro Gln 180 185 190Pro Asp Lys
Glu Pro Gly Thr Val Phe His Val Gln Gln Leu Gly Phe 195 200 205Thr
Leu Asn Asp Arg Val Ile Arg Pro Ala Lys Val Gly Ile Val Lys 210 215
220Gly Glu Glu Asn22598687DNASaccharomyces cerevisiae 98atgagagctt
tttcagcagc caccgttagg gccacaacta ggaagtcgtt catcccaatg 60gcaccaagaa
ctccttttgt gactccatca tttacaaaga atgtaggctc aatgagaaga
120atgagatttt attctgatga agccaaaagt gaagaatcca aagaaaacaa
tgaagatttg 180actgaagagc aatcagaaat caagaaatta gagagccagt
taagcgcgaa gactaaagaa 240gcttctgaac tcaaggacag attattaaga
tctgtggcag atttcagaaa tttacaacaa 300gtcacaaaga aggatattca
gaaagctaag gactttgctt tacagaagtt tgcaaaggat 360ttattggaat
ctgtagataa ctttggtcat gctttgaatg cttttaaaga ggaagactta
420caaaagtcca aggaaattag tgatttgtat acaggggtta gaatgacaag
agatgttttt 480gaaaacaccc taagaaagca cggtattgaa aaattagacc
cattgggaga accatttgat 540ccaaataaac acgaagcaac gttcgagttg
ccacaacctg ataaggaacc gggtactgtt 600ttccatgtac aacaattagg
tttcaccttg aatgacagag ttatcagacc agcaaaagtc 660ggaattgtta
agggcgaaga gaactaa 68799290PRTSaccharomyces cerevisiae 99Met Glu
Lys Leu Leu Gln Trp Ser Ile Ala Asn Ser Gln Gly Asp Lys1 5 10 15Glu
Ala Met Ala Arg Ala Gly Gln Pro Asp Pro Lys Leu Leu Gln Gln 20 25
30Leu Phe Gly Gly Gly Gly Pro Asp Asp Pro Thr Leu Met Lys Glu Ser
35 40 45Met Ala Val Ile Met Asn Pro Glu Val Asp Leu Glu Thr Lys Leu
Val 50 55 60Ala Phe Asp Asn Phe Glu Met Leu Ile Glu Asn Leu Asp Asn
Ala Asn65 70 75 80Asn Ile Glu Asn Leu Lys Leu Trp Glu Pro Leu Leu
Asp Val Leu Val 85 90 95Gln Thr Lys Asp Glu Glu Leu Arg Ala Ala Ala
Leu Ser Ile Ile Gly 100 105 110Thr Ala Val Gln Asn Asn Leu Asp Ser
Gln Asn Asn Phe Met Lys Tyr 115 120 125Asp Asn Gly Leu Arg Ser Leu
Ile Glu Ile Ala Ser Asp Lys Thr Lys 130 135 140Pro Leu Asp Val Arg
Thr Lys Ala Phe Tyr Ala Leu Ser Asn Leu Ile145 150 155 160Arg Asn
His Lys Asp Ile Ser Glu Lys Phe Phe Lys Leu Asn Gly Leu 165 170
175Asp Cys Ile Ala Pro Val Leu Ser Asp Asn Thr Ala Lys Pro Lys Leu
180 185 190Lys Met Arg Ala Ile Ala Leu Leu Thr Ala Tyr Leu Ser Ser
Val Lys 195 200 205Ile Asp Glu Asn Ile Ile Ser Val Leu Arg Lys Asp
Gly Val Ile Glu 210 215 220Ser Thr Ile Glu Cys Leu Ser Asp Glu Ser
Asn Leu Asn Ile Ile Asp225 230 235 240Arg Val Leu Ser Phe Leu Ser
His Leu Ile Ser Ser Gly Ile Lys Phe 245 250 255Asn Glu Gln Glu Leu
His Lys Leu Asn Glu Gly Tyr Lys His Ile Glu 260 265 270Pro Leu Lys
Asp Arg Leu Asn Glu Asp Asp Tyr Leu Ala Val Lys Tyr 275 280 285Val
Leu 290100873DNASaccharomyces cerevisiae 100atggaaaagc tattacagtg
gtctattgcg aattctcaag gggacaaaga agctatggct 60agggccggcc aacctgatcc
taaattgcta cagcagttat tcggtggtgg tggtcctgac 120gatccaacct
taatgaaaga atccatggct gttattatga atccggaggt tgacttagaa
180acaaaactcg ttgcatttga caactttgaa atgttgattg agaacttaga
taatgctaat 240aatatcgaaa atttaaaact gtgggagcca ttgttggatg
ttcttgttca gacgaaggat 300gaagaactac gtgctgctgc tttatccatt
attggaacgg ctgtgcaaaa caacttggat 360tcgcaaaata atttcatgaa
atacgacaat ggtctgcgaa gccttatcga aatagctagt 420gacaagacaa
agccactcga cgtgagaaca aaagcttttt acgcactatc taatctaata
480agaaaccaca aagatatctc agaaaagttt ttcaaattaa atgggctcga
ctgcatagca 540cctgtattaa gtgataacac cgccaaacca aaactgaaaa
tgagagccat tgccttattg 600accgcatatt tgtcatctgt taagattgat
gaaaatataa tcagtgtgct gagaaaggat 660ggagtaattg aaagtacgat
tgagtgcttg tctgacgaga gtaacttgaa catcatagat 720agagttctgt
cttttctctc tcacctgata tcttccggaa taaaatttaa tgaacaggaa
780ttgcacaaat tgaacgaagg ttacaaacat atcgagcctc taaaggacag
acttaatgaa 840gacgattatt tagccgtaaa gtatgtatta tga
87310140DNAArtificial SequencePrimer HO 5' ForNotIBbsI
101gcatgcggcc gcccgaagac cctacacagg gcttaagggc 4010235DNAArtificial
SequencePrimer HO 5' RevBsiWIMluI 102ccacgcgtcg tacgggattg
ctgcttatga ggata 3510337DNAArtificial SequencePrimer HO 3'
ForMluIEcoRI 103acgcgtgaat tcaaaaaggg aacccgtata tttcagc
3710439DNAArtificial SequencePrimer HO 3' RevBbsIClaI 104tatcgatagt
cttcctaata tacacatttt agcagatgc 39105102DNAArtificial
SequencePrimer pBST HO Poly For 105gcatgcatac gcgtcacgca tgtgcctcag
cggccggccg gcgccgggcc ccggaccgcc 60tgcaggctcg agttaattaa gtttaaacga
attcgcatgc at 102106102DNAArtificial SequencePrimer pBST HO Poly
Rev 106atgcatgcga attcgtttaa acttaattaa ctcgagcctg caggcggtcc
ggggcccggc 60gccggccggc cgctgaggca catgcgtgac gcgtatgcat gc
10210762DNAArtificial SequencePrimer Ycplac33 Poly For
107ctagattgga tccctagtct aggtttaaac tagcgattca cctaggtgct
aggaattcta 60gc 6210862DNAArtificial SequencePrimer Ycplac33 Poly
Rev 108gctagaattc ctagcaccta ggtgaatcgc tagtttaaac ctagactagg
gatccaatct 60ag 6210969DNAArtificial SequencePrimer LHS1forOverlap
109cacaatattt caagctatac caagcataca atcaactatc tcatatacaa
tgcgaaacgt 60tttaaggct 6911034DNAArtificial SequencePrimer
LHS1revBbvCI 110gcatgctgag ggtgccacta taatattaat gtgc
3411171DNAArtificial SequencePrimer SLS1forOverlap 111caccaacaca
cacaaaaaac agtacttcac taaatttaca cacaaaacaa aatggtccgg 60attcttccca
t 7111231DNAArtificial SequencePrimer SLS1revNarI 112gcatggcgcc
ccacggcagg gcagttggca c 3111370DNAArtificial SequencePrimer
JEM1forOverlap 113cagatcatca aggaagtaat tatctacttt ttacaacaaa
tataaaacaa tgatactgat 60ctcgggatac 7011433DNAArtificial
SequencePrimer JEM1revRsrII 114cgatcggtcc gagggaaata aggcagatca aag
3311572DNAArtificial SequencePrimer SCJ1forOverlap 115cacgcttact
gcttttttct tcccaagatc gaaaatttac tgaattaaca atgattccaa 60aattatatat
ac 7211629DNAArtificial SequencePrimer SCJ1revXhoI 116gcatctcgag
gactttgaga cctgtgatc 2911738DNAArtificial SequencePrimer
ADH1promForAleI 117cgatcaccga tgtggttgtt tccgggtgta caatatgg
3811871DNAArtificial SequencePrimer ADH1promRevOverlap
118cctatagcaa caaaagctgt taaaaataaa agccttaaaa cgtttcgcat
tgtatatgag 60atagttgatt g 7111932DNAArtificial SequencePrimer
PGK1promForPspOMI 119gcatgggccc agattcctga cttcaactca ag
3212073DNAArtificial SequencePrimer PGK1promRevOverlap
120ggcaaaataa cgctatacac taaaagacag tatcccgaga tcagtatcat
tgttttatat 60ttgttgtaaa aac 7312133DNAArtificial SequencePrimer
TDH1promForFseI 121gcatggccgg ccaccatatg gaggataagt tgg
3312271DNAArtificial SequencePrimer TDH1promRevOverlap
122ctaatttcga agatagggcg ctcaaaatta tgggaagaat ccggaccatt
ttgttttgtg 60tgttttaaat c 7112336DNAArtificial SequencePrimer
TEF1promForSbfI 123cggtagtacc tgcaggaagc aacaggcgcg ttggac
3612478DNAArtificial SequencePrimer TEF1promRevOverlap
124ggcaacaaca ataaagatag tatcaaatgt atatataatt ttggaatcat
tttgtaatta 60aaacttagat tagattgc 7812532DNAArtificial
SequencePrimer URA3forPac1 125ctagagttaa ttaagtttca attcaattca tc
3212632DNAArtificial SequencePrimer URA3revPme1 126gcctgagttt
aaacgttttc tttccaattt tt 3212740DNAArtificial SequencePrimer A01
127gcatgcggcc gcccgaagac cctacacagg gcttaagggc 4012835DNAArtificial
SequencePrimer A02 128ccacgcgtcg tacgggattg ctgcttatga ggata
3512937DNAArtificial SequencePrimer A03 129acgcgtgaat tcaaaaaggg
aacccgtata tttcagc 3713039DNAArtificial SequencePrimer A04
130tatcgatagt cttcctaata tacacatttt agcagatgc 39131102DNAArtificial
SequencePrimer A05 131gcatgcatac gcgtcacgca tgtgcctcag cggccggccg
gcgccgggcc ccggaccgcc 60tgcaggctcg agttaattaa gtttaaacga attcgcatgc
at 102132102DNAArtificial SequencePrimer A06 132atgcatgcga
attcgtttaa acttaattaa ctcgagcctg caggcggtcc ggggcccggc 60gccggccggc
cgctgaggca catgcgtgac gcgtatgcat gc 10213335DNAArtificial
SequencePrimer A07 133ctaggtaact taattaaggg taagctgcca cagca
3513434DNAArtificial SequencePrimer A08 134ctacgtactc tagatgttaa
ttcagtaaat tttc 3413530DNAArtificial SequencePrimer A09
135ctagactcta gatctctgct tttgtgcgcg 3013634DNAArtificial
SequencePrimer A10 136catgctacgt ttaaacgatg atcatatgat acac
3413732DNAArtificial SequencePrimer A11 137ctagagttaa ttaagtttca
attcaattca tc 3213832DNAArtificial SequencePrimer A12 138gcctgagttt
aaacgttttc tttccaattt tt 3213962DNAArtificial SequencePrimer A13
139ctagattgga tccctagtct aggtttaaac tagcgattca cctaggtgct
aggaattcta 60gc 6214062DNAArtificial SequencePrimer A14
140gctagaattc ctagcaccta ggtgaatcgc tagtttaaac ctagactagg
gatccaatct 60ag 6214169DNAArtificial SequencePrimer C01
141cacaatattt caagctatac caagcataca atcaactatc tcatatacaa
tgcgaaacgt 60tttaaggct 6914234DNAArtificial SequencePrimer C02
142gcatgctgag ggtgccacta taatattaat gtgc 3414330DNAArtificial
SequencePrimer C03 143ctagatctct agaatggtcc ggattcttcc
3014431DNAArtificial SequencePrimer C04 144gcatggcgcc ccacggcagg
gcagttggca c 3114530DNAArtificial SequencePrimer C05 145ctagatctct
agaatgatac tgatctcggg 3014633DNAArtificial SequencePrimer C06
146cgatcggtcc gagggaaata aggcagatca aag 3314772DNAArtificial
SequencePrimer C07 147cacgcttact gcttttttct tcccaagatc gaaaatttac
tgaattaaca atgattccaa 60aattatatat ac 7214829DNAArtificial
SequencePrimer C08 148gcatctcgag gactttgaga cctgtgatc
2914938DNAArtificial SequencePrimer C09 149cgatcaccga tgtggttgtt
tccgggtgta caatatgg 3815071DNAArtificial SequencePrimer C10
150cctatagcaa caaaagctgt taaaaataaa agccttaaaa cgtttcgcat
tgtatatgag 60atagttgatt g 7115132DNAArtificial SequencePrimer C11
151gcatgggccc agattcctga cttcaactca ag 3215232DNAArtificial
SequencePrimer C12 152gatctagtct agatgtttta tatttgttgt aa
3215333DNAArtificial SequencePrimer C13 153gcatggccgg ccaccatatg
gaggataagt tgg 3315434DNAArtificial SequencePrimer C14
154acctagtcta gatttgtttt gtgtgtaaat ttag 3415536DNAArtificial
SequencePrimer C15 155cggtagtacc tgcaggaagc aacaggcgcg ttggac
3615678DNAArtificial SequencePrimer C16 156ggcaacaaca ataaagatag
tatcaaatgt atatataatt ttggaatcat tttgtaatta 60aaacttagat tagattgc
7815734DNAArtificial SequencePrimer C17 157ctagtctcta gaatggaaat
gactgatttt gaac 3415831DNAArtificial SequencePrimer C18
158ctagtctaga tcatgaagtg atgaagaaat c 3115923DNAArtificial
SequenceACT1 Forward Primer 159cccagaagct ttgttccatc ctt
2316028DNAArtificial SequenceACT1 Reverse Primer 160atgatggagt
tgtaagtagt ttggtcaa 2816120DNAArtificial SequenceACT1 Probe
161cagattccaa acccaaaaca 2016225DNAArtificial SequenceLHS1 Forward
Primer 162acactactca gcccgttaca ataga 2516325DNAArtificial
SequenceLHS1 Reverse Primer 163gtaaactttg caccacctag atgtg
2516423DNAArtificial SequenceLHS1 Probe 164atttgaagga tatgggtata
atc 2316529DNAArtificial SequenceSIL1 Forward Primer 165gacatgtacg
aaaatgacga tacaaatct 2916619DNAArtificial SequenceSIL1 Reverse
Primer 166tcgtttgccc actcttgca
1916717DNAArtificial SequenceSIL1 Probe 167tttgacgacc aattctc
1716817DNAArtificial SequenceSCJ1 Forward Primer 168ggcgcaggtg
gattcca 1716918DNAArtificial SequenceSCJ1 Reverse Primer
169cgccaggacc tccatgac 1817020DNAArtificial SequenceSCJ1 Probe
170catattcgaa cggatgtttc 2017118DNAArtificial SequenceJEM1 Forward
Primer 171cctctccacg cacatcga 1817225DNAArtificial SequenceJEM1
Reverse Primer 172tgcttgtcga ggattgtttc gtaat 2517319DNAArtificial
SequenceJEM1 Probe 173tcgttagctg ctgctatca 1917419DNAArtificial
SequenceHAC1 Forward Primer 174gaagacgcgt tgacttgca
1917525DNAArtificial SequenceHAC1 Reverse Primer 175gaaatccctg
tactcgtcaa gagaa 2517618DNAArtificial SequenceHAC1 Probe
176ccacgacgct tttgttgc 1817726DNAArtificial SequenceHAC1i Forward
Primer 177acaattcaat tgatcttgac aattgg 2617825DNAArtificial
SequenceHAC1i Reverse Primer 178tcaattcaaa tgaatcaaac ctgac
2517917DNAArtificial SequenceHAC1i Probe 179cgtaatccag aagcgca
17180157DNAArtificial SequenceOligonucleotide linker sense for
construction of pDB2283 180gagtccaatt agcttcatcg ccaataaaaa
aacaagctaa acctaattct aacaagcaaa 60gatgaagtgg gtaagcttaa cctaattcta
acaagcaaag atgcttttgc aagccttcct 120tttccttttg gctggttttg
cagccaaaat atctgca 157181161DNAArtificial SequenceSingle stranded
oligonucleotide 1 for pDB2283 181ttaagagtcc aattagcttc atcgccaata
aaaaaacaag ctaaacctaa ttctaacaag 60caaagatgaa gtgggtaagc ttaacctaat
tctaacaagc aaagatgctt ttgcaagcct 120tccttttcct tttggctggt
tttgcagcca aaatatctgc a 161182157DNAArtificial SequenceSingle
stranded oligonucleotide 2 for pDB2283 182tgcagatatt ttggctgcaa
aaccagccaa aaggaaaagg aaggcttgca aaagcatctt 60tgcttgttag aattaggtta
agcttaccca cttcatcttt gcttgttaga attaggttta 120gcttgttttt
ttattggcga tgaagctaat tggactc 157183320DNAArtificial
SequenceSynthetic phosphorylated oligonucleotide linker sense for
construction of pDB2284 183ttaagagtcc aattagcttc atcgccaata
aaaaaacaaa ctaaacctaa ttctaacaag 60caaagatgag atttccttca atttttactg
cagttttatt cgcagcatcc tccgcattag 120ctgctccagt caacactaca
acagaagatg aaacggcaca aattccggct gaagctgtca 180tcggttactc
agatttagaa ggggatttcg atgttgctgt tttgccattt tccaacagca
240caaataacgg gttattgttt ataaatacta ctattgccag cattgctgct
aaagaagaag 300gggtaagctt ggataaaaga 320184316DNAArtificial
SequenceSynthetic phosphorylated oligonucleotide linker anti sense
(pDB2283) 184tcttttatcc aagcttaccc cttcttcttt agcagcaatg ctggcaatag
tagtatttat 60aaacaataac ccgttatttg tgctgttgga aaatggcaaa acagcaacat
cgaaatcccc 120ttctaaatct gagtaaccga tgacagcttc agccggaatt
tgtgccgttt catcttctgt 180tgtagtgttg actggagcag ctaatgcgga
ggatgctgcg aataaaactg cagtaaaaat 240tgaaggaaat ctcatctttg
cttgttagaa ttaggtttag tttgtttttt tattggcgat 300gaagctaatt ggactc
31618585PRTSaccharomyces cerevisiae 185Met Arg Phe Pro Ser Ile Phe
Thr Ala Val Leu Phe Ala Ala Ser Ser1 5 10 15Ala Leu Ala Ala Pro Val
Asn Thr Thr Thr Glu Asp Glu Thr Ala Gln 20 25 30Ile Pro Ala Glu Ala
Val Ile Gly Tyr Ser Asp Leu Glu Gly Asp Phe 35 40 45Asp Val Ala Val
Leu Pro Phe Ser Asn Ser Thr Asn Asn Gly Leu Leu 50 55 60Phe Ile Asn
Thr Thr Ile Ala Ser Ile Ala Ala Lys Glu Glu Gly Val65 70 75 80Ser
Leu Asp Lys Arg 8518698DNAArtificial SequenceComplimentary SS oligo
1 186ttaagagtcc aattagcttc atcgccaata aaaaaacaaa ctaaacctaa
ttctaacaag 60caaagatgag atttccttca atttttactg cagtttta
98187110DNAArtificial SequenceComplimentary SS oligo 2
187ttcgcagcat cctccgcatt agctgctcca gtcaacacta caacagaaga
tgaaacggca 60caaattccgg ctgaagctgt catcggttac tcagatttag aaggggattt
110188112DNAArtificial SequenceComplimentary SS oligo 3
188cgatgttgct gttttgccat tttccaacag cacaaataac gggttattgt
ttataaatac 60tactattgcc agcattgctg ctaaagaaga aggggtaagc ttggataaaa
ga 112189102DNAArtificial SequenceComplimentary SS oligo 4
189tcttttatcc aagcttaccc cttcttcttt agcagcaatg ctggcaatag
tagtatttat 60aaacaataac ccgttatttg tgctgttgga aaatggcaaa ac
102190110DNAArtificial SequenceComplimentary SS oligo 5
190agcaacatcg aaatcccctt ctaaatctga gtaaccgatg acagcttcag
ccggaatttg 60tgccgtttca tcttctgttg tagtgttgac tggagcagct aatgcggagg
110191104DNAArtificial SequenceComplimentary SS oligo 6
191atgctgcgaa taaaactgca gtaaaaattg aaggaaatct catctttgct
tgttagaatt 60aggtttagtt tgttttttta ttggcgatga agctaattgg actc
10419277DNAArtificial SequenceSINK1 192gtaccaagct ttatttccct
tctttttctc tttagctcgg cttattccag gagcttggat 60aaaagagcac ccgcccg
7719378DNAArtificial SequenceSINK2 193gtgaccgggc gggtgctctt
ttatccaagc tcctggaata agccgagcta aagagaaaaa 60gaagggaaat aaagcttg
78194450DNAArtificial SequenceHuman GMCSF with incorporated
N-terminal Met codon 194aagcttacct gccatggcac ccgcccggtc acccagcccc
agcacgcagc cctgggagca 60tgtgaatgcc atccaggagg cccggcgtct cctgaacctg
agtagagaca ctgctgctga 120gatgaatgaa acagtagaag tgatatcaga
aatgtttgac ctccaggagc cgacttgcct 180acagacccgc ctggagctgt
acaagcaggg cctgcggggc agcctcacca agctcaaggg 240ccccttgacc
atgatggcca gccactacaa gcagcactgc cctccaaccc cggaaacttc
300ctgtgcaacc cagattatca cctttgaaag tttcaaagag aacctgaagg
acttcctgct 360tgtcatcccc tttgactgct gggagccagt ccaggagtga
taaggatccg aattcgtaat 420catggtcata gctgtttcct gtgtgaaatt
450195128PRTArtificial SequenceHuman GMCSF with incorporated
N-terminal Met codon 195Met Ala Pro Ala Arg Ser Pro Ser Pro Ser Thr
Gln Pro Trp Glu His1 5 10 15Val Asn Ala Ile Gln Glu Ala Arg Arg Leu
Leu Asn Leu Ser Arg Asp 20 25 30Thr Ala Ala Glu Met Asn Glu Thr Val
Glu Val Ile Ser Glu Met Phe 35 40 45Asp Leu Gln Glu Pro Thr Cys Leu
Gln Thr Arg Leu Glu Leu Tyr Lys 50 55 60Gln Gly Leu Arg Gly Ser Leu
Thr Lys Leu Lys Gly Pro Leu Thr Met65 70 75 80Met Ala Ser His Tyr
Lys Gln His Cys Pro Pro Thr Pro Glu Thr Ser 85 90 95Cys Ala Thr Gln
Ile Ile Thr Phe Glu Ser Phe Lys Glu Asn Leu Lys 100 105 110Asp Phe
Leu Leu Val Ile Pro Phe Asp Cys Trp Glu Pro Val Gln Glu 115 120
125
* * * * *
References